Full text data of OTUB1
OTUB1
(OTB1, OTU1)
[Confidence: low (only semi-automatic identification from reviews)]
Ubiquitin thioesterase OTUB1; 3.4.19.12 (Deubiquitinating enzyme OTUB1; OTU domain-containing ubiquitin aldehyde-binding protein 1; Otubain-1; hOTU1; Ubiquitin-specific-processing protease OTUB1)
Note: presumably soluble (membrane word is not in UniProt keywords or features)
Ubiquitin thioesterase OTUB1; 3.4.19.12 (Deubiquitinating enzyme OTUB1; OTU domain-containing ubiquitin aldehyde-binding protein 1; Otubain-1; hOTU1; Ubiquitin-specific-processing protease OTUB1)
Note: presumably soluble (membrane word is not in UniProt keywords or features)
UniProt
Q96FW1
ID OTUB1_HUMAN Reviewed; 271 AA.
AC Q96FW1; Q32Q78; Q96II3; Q9NXQ4; Q9P0B8;
DT 01-MAR-2004, integrated into UniProtKB/Swiss-Prot.
read moreDT 01-MAR-2004, sequence version 2.
DT 22-JAN-2014, entry version 115.
DE RecName: Full=Ubiquitin thioesterase OTUB1;
DE EC=3.4.19.12;
DE AltName: Full=Deubiquitinating enzyme OTUB1;
DE AltName: Full=OTU domain-containing ubiquitin aldehyde-binding protein 1;
DE AltName: Full=Otubain-1;
DE Short=hOTU1;
DE AltName: Full=Ubiquitin-specific-processing protease OTUB1;
GN Name=OTUB1; Synonyms=OTB1, OTU1; ORFNames=HSPC263;
OS Homo sapiens (Human).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;
OC Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini;
OC Catarrhini; Hominidae; Homo.
OX NCBI_TaxID=9606;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORM 1), IDENTIFICATION BY MASS
RP SPECTROMETRY, FUNCTION, TISSUE SPECIFICITY, AND MUTAGENESIS OF ASP-88;
RP CYS-91 AND HIS-265.
RC TISSUE=Cervix carcinoma;
RX PubMed=12704427; DOI=10.1038/sj.embor.embor824;
RA Balakirev M.Y., Tcherniuk S.O., Jaquinod M., Chroboczek J.;
RT "Otubains: a new family of cysteine proteases in the ubiquitin
RT pathway.";
RL EMBO Rep. 4:517-522(2003).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS 1 AND 2), ALTERNATIVE SPLICING,
RP FUNCTION, TISSUE SPECIFICITY, INTERACTION WITH RNF128 AND USP8, AND
RP MUTAGENESIS OF CYS-91; ARG-176 AND CYS-212.
RX PubMed=14661020; DOI=10.1038/ni1017;
RA Soares L., Seroogy C., Skrenta H., Anandasabapathy N., Lovelace P.,
RA Chung C.D., Engleman E., Fathman C.G.;
RT "Two isoforms of otubain 1 regulate T cell anergy via GRAIL.";
RL Nat. Immunol. 5:45-54(2004).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM 1).
RC TISSUE=Umbilical cord blood;
RX PubMed=11042152; DOI=10.1101/gr.140200;
RA Zhang Q.-H., Ye M., Wu X.-Y., Ren S.-X., Zhao M., Zhao C.-J., Fu G.,
RA Shen Y., Fan H.-Y., Lu G., Zhong M., Xu X.-R., Han Z.-G., Zhang J.-W.,
RA Tao J., Huang Q.-H., Zhou J., Hu G.-X., Gu J., Chen S.-J., Chen Z.;
RT "Cloning and functional analysis of cDNAs with open reading frames for
RT 300 previously undefined genes expressed in CD34+ hematopoietic
RT stem/progenitor cells.";
RL Genome Res. 10:1546-1560(2000).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM 1).
RC TISSUE=Colon;
RX PubMed=14702039; DOI=10.1038/ng1285;
RA Ota T., Suzuki Y., Nishikawa T., Otsuki T., Sugiyama T., Irie R.,
RA Wakamatsu A., Hayashi K., Sato H., Nagai K., Kimura K., Makita H.,
RA Sekine M., Obayashi M., Nishi T., Shibahara T., Tanaka T., Ishii S.,
RA Yamamoto J., Saito K., Kawai Y., Isono Y., Nakamura Y., Nagahari K.,
RA Murakami K., Yasuda T., Iwayanagi T., Wagatsuma M., Shiratori A.,
RA Sudo H., Hosoiri T., Kaku Y., Kodaira H., Kondo H., Sugawara M.,
RA Takahashi M., Kanda K., Yokoi T., Furuya T., Kikkawa E., Omura Y.,
RA Abe K., Kamihara K., Katsuta N., Sato K., Tanikawa M., Yamazaki M.,
RA Ninomiya K., Ishibashi T., Yamashita H., Murakawa K., Fujimori K.,
RA Tanai H., Kimata M., Watanabe M., Hiraoka S., Chiba Y., Ishida S.,
RA Ono Y., Takiguchi S., Watanabe S., Yosida M., Hotuta T., Kusano J.,
RA Kanehori K., Takahashi-Fujii A., Hara H., Tanase T.-O., Nomura Y.,
RA Togiya S., Komai F., Hara R., Takeuchi K., Arita M., Imose N.,
RA Musashino K., Yuuki H., Oshima A., Sasaki N., Aotsuka S.,
RA Yoshikawa Y., Matsunawa H., Ichihara T., Shiohata N., Sano S.,
RA Moriya S., Momiyama H., Satoh N., Takami S., Terashima Y., Suzuki O.,
RA Nakagawa S., Senoh A., Mizoguchi H., Goto Y., Shimizu F., Wakebe H.,
RA Hishigaki H., Watanabe T., Sugiyama A., Takemoto M., Kawakami B.,
RA Yamazaki M., Watanabe K., Kumagai A., Itakura S., Fukuzumi Y.,
RA Fujimori Y., Komiyama M., Tashiro H., Tanigami A., Fujiwara T.,
RA Ono T., Yamada K., Fujii Y., Ozaki K., Hirao M., Ohmori Y.,
RA Kawabata A., Hikiji T., Kobatake N., Inagaki H., Ikema Y., Okamoto S.,
RA Okitani R., Kawakami T., Noguchi S., Itoh T., Shigeta K., Senba T.,
RA Matsumura K., Nakajima Y., Mizuno T., Morinaga M., Sasaki M.,
RA Togashi T., Oyama M., Hata H., Watanabe M., Komatsu T.,
RA Mizushima-Sugano J., Satoh T., Shirai Y., Takahashi Y., Nakagawa K.,
RA Okumura K., Nagase T., Nomura N., Kikuchi H., Masuho Y., Yamashita R.,
RA Nakai K., Yada T., Nakamura Y., Ohara O., Isogai T., Sugano S.;
RT "Complete sequencing and characterization of 21,243 full-length human
RT cDNAs.";
RL Nat. Genet. 36:40-45(2004).
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RX PubMed=16554811; DOI=10.1038/nature04632;
RA Taylor T.D., Noguchi H., Totoki Y., Toyoda A., Kuroki Y., Dewar K.,
RA Lloyd C., Itoh T., Takeda T., Kim D.-W., She X., Barlow K.F.,
RA Bloom T., Bruford E., Chang J.L., Cuomo C.A., Eichler E.,
RA FitzGerald M.G., Jaffe D.B., LaButti K., Nicol R., Park H.-S.,
RA Seaman C., Sougnez C., Yang X., Zimmer A.R., Zody M.C., Birren B.W.,
RA Nusbaum C., Fujiyama A., Hattori M., Rogers J., Lander E.S.,
RA Sakaki Y.;
RT "Human chromosome 11 DNA sequence and analysis including novel gene
RT identification.";
RL Nature 440:497-500(2006).
RN [6]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM 1).
RC TISSUE=Colon, and Skin;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA
RT project: the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [7]
RP PARTIAL PROTEIN SEQUENCE, AND FUNCTION.
RX PubMed=12401499; DOI=10.1016/S1074-5521(02)00248-X;
RA Borodovsky A., Ovaa H., Kolli N., Gan-Erdene T., Wilkinson K.D.,
RA Ploegh H.L., Kessler B.M.;
RT "Chemistry-based functional proteomics reveals novel members of the
RT deubiquitinating enzyme family.";
RL Chem. Biol. 9:1149-1159(2002).
RN [8]
RP PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-16, AND MASS
RP SPECTROMETRY.
RC TISSUE=Cervix carcinoma;
RX PubMed=18669648; DOI=10.1073/pnas.0805139105;
RA Dephoure N., Zhou C., Villen J., Beausoleil S.A., Bakalarski C.E.,
RA Elledge S.J., Gygi S.P.;
RT "A quantitative atlas of mitotic phosphorylation.";
RL Proc. Natl. Acad. Sci. U.S.A. 105:10762-10767(2008).
RN [9]
RP ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, MASS SPECTROMETRY, AND
RP CLEAVAGE OF INITIATOR METHIONINE.
RX PubMed=19413330; DOI=10.1021/ac9004309;
RA Gauci S., Helbig A.O., Slijper M., Krijgsveld J., Heck A.J.,
RA Mohammed S.;
RT "Lys-N and trypsin cover complementary parts of the phosphoproteome in
RT a refined SCX-based approach.";
RL Anal. Chem. 81:4493-4501(2009).
RN [10]
RP FUNCTION, INTERACTION WITH ESR1, AND MUTAGENESIS OF CYS-91.
RX PubMed=19383985; DOI=10.1074/jbc.M109.007484;
RA Stanisic V., Malovannaya A., Qin J., Lonard D.M., O'Malley B.W.;
RT "OTU Domain-containing ubiquitin aldehyde-binding protein 1 (OTUB1)
RT deubiquitinates estrogen receptor (ER) alpha and affects ERalpha
RT transcriptional activity.";
RL J. Biol. Chem. 284:16135-16145(2009).
RN [11]
RP FUNCTION, UBIQUITIN-BINDING, DOMAIN, AND MUTAGENESIS OF CYS-23; CYS-91
RP AND CYS-212.
RX PubMed=19211026; DOI=10.1016/j.jmb.2008.12.085;
RA Wang T., Yin L., Cooper E.M., Lai M.-Y., Dickey S., Pickart C.M.,
RA Fushman D., Wilkinson K.D., Cohen R.E., Wolberger C.;
RT "Evidence for bidentate substrate binding as the basis for the K48
RT linkage specificity of otubain 1.";
RL J. Mol. Biol. 386:1011-1023(2009).
RN [12]
RP FUNCTION IN INHIBITION OF RNF168, INTERACTION WITH UBE2N/UBC13, AND
RP MUTAGENESIS OF ASP-88; CYS-91 AND HIS-265.
RX PubMed=20725033; DOI=10.1038/nature09297;
RA Nakada S., Tai I., Panier S., Al-Hakim A., Iemura S., Juang Y.C.,
RA O'Donnell L., Kumakubo A., Munro M., Sicheri F., Gingras A.C.,
RA Natsume T., Suda T., Durocher D.;
RT "Non-canonical inhibition of DNA damage-dependent ubiquitination by
RT OTUB1.";
RL Nature 466:941-946(2010).
RN [13]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX PubMed=21269460; DOI=10.1186/1752-0509-5-17;
RA Burkard T.R., Planyavsky M., Kaupe I., Breitwieser F.P.,
RA Buerckstuemmer T., Bennett K.L., Superti-Furga G., Colinge J.;
RT "Initial characterization of the human central proteome.";
RL BMC Syst. Biol. 5:17-17(2011).
RN [14]
RP ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, AND MASS SPECTROMETRY.
RX PubMed=22814378; DOI=10.1073/pnas.1210303109;
RA Van Damme P., Lasa M., Polevoda B., Gazquez C., Elosegui-Artola A.,
RA Kim D.S., De Juan-Pardo E., Demeyer K., Hole K., Larrea E.,
RA Timmerman E., Prieto J., Arnesen T., Sherman F., Gevaert K.,
RA Aldabe R.;
RT "N-terminal acetylome analyses and functional insights of the N-
RT terminal acetyltransferase NatB.";
RL Proc. Natl. Acad. Sci. U.S.A. 109:12449-12454(2012).
RN [15]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=23827681; DOI=10.1016/j.cell.2013.05.046;
RA Mevissen T.E., Hospenthal M.K., Geurink P.P., Elliott P.R., Akutsu M.,
RA Arnaudo N., Ekkebus R., Kulathu Y., Wauer T., El Oualid F.,
RA Freund S.M., Ovaa H., Komander D.;
RT "OTU deubiquitinases reveal mechanisms of linkage specificity and
RT enable ubiquitin chain restriction analysis.";
RL Cell 154:169-184(2013).
RN [16]
RP X-RAY CRYSTALLOGRAPHY (1.69 ANGSTROMS) OF 40-271, FUNCTION,
RP INTERACTION WITH FUS AND GNB2L1, AND MUTAGENESIS OF PRO-87 AND CYS-91.
RX PubMed=18954305; DOI=10.1042/BJ20081318;
RA Edelmann M.J., Iphoefer A., Akutsu M., Altun M., di Gleria K.,
RA Kramer H.B., Fiebiger E., Dhe-Paganon S., Kessler B.M.;
RT "Structural basis and specificity of human otubain 1-mediated
RT deubiquitination.";
RL Biochem. J. 418:379-390(2009).
RN [17]
RP X-RAY CRYSTALLOGRAPHY (3.3 ANGSTROMS) OF 25-271 IN COMPLEX WITH UBE2V2
RP AND UBE2N, FREE UBIQUITIN-BINDING, ENZYME REGULATION, AND MUTAGENESIS
RP OF GLN-33; ILE-37; GLN-39; CYS-91; ALA-116; THR-134; ASP-137; PHE-190;
RP TYR-261 AND PRO-263.
RX PubMed=22325355; DOI=10.1016/j.molcel.2012.01.011;
RA Juang Y.C., Landry M.C., Sanches M., Vittal V., Leung C.C.,
RA Ceccarelli D.F., Mateo A.R., Pruneda J.N., Mao D.Y., Szilard R.K.,
RA Orlicky S., Munro M., Brzovic P.S., Klevit R.E., Sicheri F.,
RA Durocher D.;
RT "OTUB1 co-opts Lys48-linked ubiquitin recognition to suppress E2
RT enzyme function.";
RL Mol. Cell 45:384-397(2012).
RN [18]
RP X-RAY CRYSTALLOGRAPHY (3.11 ANGSTROMS) OF 1-45 IN COMPLEX WITH UBE2N
RP AND UBIQUITIN, ENZYME REGULATION, AND FREE UBIQUITIN-BINDING.
RX PubMed=22367539; DOI=10.1038/nature10911;
RA Wiener R., Zhang X., Wang T., Wolberger C.;
RT "The mechanism of OTUB1-mediated inhibition of ubiquitination.";
RL Nature 483:618-622(2012).
CC -!- FUNCTION: Hydrolase that can specifically remove 'Lys-48'-linked
CC conjugated ubiquitin from proteins and plays an important
CC regulatory role at the level of protein turnover by preventing
CC degradation. Regulator of T-cell anergy, a phenomenon that occurs
CC when T-cells are rendered unresponsive to antigen rechallenge and
CC no longer respond to their cognate antigen. Acts via its
CC interaction with RNF128/GRAIL, a crucial inductor of CD4 T-cell
CC anergy. Isoform 1 destabilizes RNF128, leading to prevent anergy.
CC In contrast, isoform 2 stabilizes RNF128 and promotes anergy.
CC Surprisingly, it regulates RNF128-mediated ubiquitination, but
CC does not deubiquitinate polyubiquitinated RNF128. Deubiquitinates
CC estrogen receptor alpha (ESR1). Mediates deubiquitination of 'Lys-
CC 48'-linked polyubiquitin chains, but not 'Lys-63'-linked
CC polyubiquitin chains. Not able to cleave di-ubiquitin. Also
CC capable of removing NEDD8 from NEDD8 conjugates, but with a much
CC lower preference compared to 'Lys-48'-linked ubiquitin.
CC -!- FUNCTION: Plays a key non-catalytic role in DNA repair regulation
CC by inhibiting activity of RNF168, an E3 ubiquitin-protein ligase
CC that promotes accumulation of 'Lys-63'-linked histone H2A and H2AX
CC at DNA damage sites. Inhibits RNF168 independently of ubiquitin
CC thioesterase activity by binding and inhibiting UBE2N/UBC13, the
CC E2 partner of RNF168, thereby limiting spreading of 'Lys-63'-
CC linked histone H2A and H2AX marks. Inhibition occurs by binding to
CC free ubiquitin: free ubiquitin acts as an allosteric regulator
CC that increases affinity for UBE2N/UBC13 and disrupts interaction
CC with UBE2V1. The OTUB1-UBE2N/UBC13-free ubiquitin complex adopts a
CC configuration that mimics a cleaved 'Lys48'-linked di-ubiquitin
CC chain.
CC -!- CATALYTIC ACTIVITY: Thiol-dependent hydrolysis of ester,
CC thioester, amide, peptide and isopeptide bonds formed by the C-
CC terminal Gly of ubiquitin (a 76-residue protein attached to
CC proteins as an intracellular targeting signal).
CC -!- ENZYME REGULATION: By free ubiquitin: binding of free ubiquitin
CC triggers conformational changes in the OTU domain and formation of
CC a ubiquitin-binding helix in the N-terminus, promoting binding of
CC the conjugated donor ubiquitin in UBE2N/UBC13 to OTUB1.
CC -!- SUBUNIT: Isoform 1 and isoform 2 interact with RNF128. Isoform 1
CC forms a ternary complex with RNF128 and USP8. Isoform 1 interacts
CC with the C-terminal UCH catalytic domain of USP8. Isoform 2 does
CC not associate with USP8. Interacts with FUS, ESR1 and
CC GNB2L1/RACK1. Interacts with UBE2N/UBC13.
CC -!- INTERACTION:
CC Q13490:BIRC2; NbExp=3; IntAct=EBI-1058491, EBI-514538;
CC Q00987:MDM2; NbExp=5; IntAct=EBI-1058491, EBI-389668;
CC P04637:TP53; NbExp=8; IntAct=EBI-1058491, EBI-366083;
CC P51668:UBE2D1; NbExp=4; IntAct=EBI-1058491, EBI-743540;
CC P62837:UBE2D2; NbExp=5; IntAct=EBI-1058491, EBI-347677;
CC P61077:UBE2D3; NbExp=4; IntAct=EBI-1058491, EBI-348268;
CC Q56921:ypkA (xeno); NbExp=5; IntAct=EBI-1058491, EBI-8022937;
CC Q9RI12:ypkA (xeno); NbExp=3; IntAct=EBI-1058491, EBI-2849107;
CC -!- SUBCELLULAR LOCATION: Cytoplasm (By similarity).
CC -!- ALTERNATIVE PRODUCTS:
CC Event=Alternative splicing; Named isoforms=2;
CC Name=1; Synonyms=Otubain-1;
CC IsoId=Q96FW1-1; Sequence=Displayed;
CC Name=2; Synonyms=ARF-1;
CC IsoId=Q96FW1-2; Sequence=VSP_009464;
CC Note=Lacks the catalytic sites for protease activity;
CC -!- TISSUE SPECIFICITY: Isoform 1 is ubiquitous. Isoform 2 is
CC expressed only in lymphoid tissues such as tonsils, lymph nodes
CC and spleen, as well as peripheral blood mononuclear cells.
CC -!- DOMAIN: In addition to ubiquitin-binding at the Cys-91 active
CC site, a proximal ubiquitin-binding site is also present at Cys-23
CC Occupancy of the active site is needed to enable tight binding to
CC the second site. Distinct binding sites for the ubiquitins may
CC allow to discriminate among different isopeptide linkages (i.e.
CC 'Lys-48'-, 'Lys-63'-linked polyubiquitin) in polyubiquitin
CC substrates and achieve linkage-specific deubiquitination.
CC -!- MISCELLANEOUS: In the structure described by PubMed:18954305, the
CC His-265 active site of the catalytic triad is located too far to
CC interact directly with the active site Cys-91. A possible
CC explanation is that OTUB1 is in inactive conformation in absence
CC of ubiquitin and a conformation change may move His-265 in the
CC proximity of Cys-91 in presence of ubiquitin substrate.
CC -!- SIMILARITY: Belongs to the peptidase C65 family.
CC -!- SIMILARITY: Contains 1 OTU domain.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAF28941.1; Type=Erroneous initiation;
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DR EMBL; AY177200; AAO27702.1; -; mRNA.
DR EMBL; AF161381; AAF28941.1; ALT_INIT; mRNA.
DR EMBL; AK000120; BAA90956.1; -; mRNA.
DR EMBL; AP000721; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR EMBL; BC007519; AAH07519.1; -; mRNA.
DR EMBL; BC010368; AAH10368.1; -; mRNA.
DR EMBL; BC107701; AAI07702.1; -; mRNA.
DR RefSeq; NP_060140.2; NM_017670.2.
DR UniGene; Hs.473788; -.
DR PDB; 2ZFY; X-ray; 1.69 A; A=40-271.
DR PDB; 3VON; X-ray; 3.15 A; A/H/O/V/c/j=45-271.
DR PDB; 4DDG; X-ray; 3.30 A; A/B/C/J/K/L=25-271.
DR PDB; 4DDI; X-ray; 3.80 A; A/B/C=25-271.
DR PDB; 4DHZ; X-ray; 3.11 A; A=1-45.
DR PDB; 4I6L; X-ray; 2.49 A; A=45-271.
DR PDB; 4LDT; X-ray; 1.90 A; A=1-45.
DR PDBsum; 2ZFY; -.
DR PDBsum; 3VON; -.
DR PDBsum; 4DDG; -.
DR PDBsum; 4DDI; -.
DR PDBsum; 4DHZ; -.
DR PDBsum; 4I6L; -.
DR PDBsum; 4LDT; -.
DR ProteinModelPortal; Q96FW1; -.
DR SMR; Q96FW1; 25-271.
DR DIP; DIP-41158N; -.
DR IntAct; Q96FW1; 26.
DR MINT; MINT-5001867; -.
DR STRING; 9606.ENSP00000301453; -.
DR MEROPS; C65.001; -.
DR PhosphoSite; Q96FW1; -.
DR DMDM; 44888286; -.
DR PaxDb; Q96FW1; -.
DR PRIDE; Q96FW1; -.
DR DNASU; 55611; -.
DR Ensembl; ENST00000428192; ENSP00000402551; ENSG00000167770.
DR Ensembl; ENST00000538426; ENSP00000444357; ENSG00000167770.
DR GeneID; 55611; -.
DR KEGG; hsa:55611; -.
DR UCSC; uc001nyf.1; human.
DR CTD; 55611; -.
DR GeneCards; GC11P063753; -.
DR HGNC; HGNC:23077; OTUB1.
DR HPA; HPA039176; -.
DR MIM; 608337; gene.
DR neXtProt; NX_Q96FW1; -.
DR PharmGKB; PA134988141; -.
DR eggNOG; NOG267426; -.
DR HOVERGEN; HBG053383; -.
DR InParanoid; Q96FW1; -.
DR KO; K09602; -.
DR PhylomeDB; Q96FW1; -.
DR ChiTaRS; OTUB1; human.
DR EvolutionaryTrace; Q96FW1; -.
DR GeneWiki; OTUB1; -.
DR GenomeRNAi; 55611; -.
DR NextBio; 60179; -.
DR PRO; PR:Q96FW1; -.
DR ArrayExpress; Q96FW1; -.
DR Bgee; Q96FW1; -.
DR CleanEx; HS_OTUB1; -.
DR Genevestigator; Q96FW1; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0004843; F:deubiquitinase activity; IDA:UniProtKB.
DR GO; GO:0019784; F:NEDD8-specific protease activity; IDA:UniProtKB.
DR GO; GO:0008242; F:omega peptidase activity; IEA:InterPro.
DR GO; GO:0043130; F:ubiquitin binding; IDA:UniProtKB.
DR GO; GO:0006974; P:cellular response to DNA damage stimulus; IDA:UniProtKB.
DR GO; GO:0006281; P:DNA repair; IEA:UniProtKB-KW.
DR GO; GO:2000780; P:negative regulation of double-strand break repair; IMP:UniProtKB.
DR GO; GO:1901315; P:negative regulation of histone H2A K63-linked ubiquitination; IDA:UniProtKB.
DR GO; GO:0071108; P:protein K48-linked deubiquitination; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR InterPro; IPR003323; OTU.
DR InterPro; IPR019400; Peptidase_C65_otubain.
DR InterPro; IPR016615; Ubiquitin_thioesterase_Otubain.
DR Pfam; PF10275; Peptidase_C65; 1.
DR PIRSF; PIRSF013503; Ubiquitin_thioesterase_Otubain; 1.
DR PROSITE; PS50802; OTU; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylation; Adaptive immunity; Alternative splicing;
KW Complete proteome; Cytoplasm; Direct protein sequencing; DNA damage;
KW DNA repair; Hydrolase; Immunity; Phosphoprotein; Protease;
KW Reference proteome; Thiol protease; Ubl conjugation pathway.
FT INIT_MET 1 1 Removed.
FT CHAIN 2 271 Ubiquitin thioesterase OTUB1.
FT /FTId=PRO_0000221008.
FT DOMAIN 80 271 OTU.
FT REGION 130 138 Ubiquitin-conjugating enzyme E2 binding.
FT REGION 169 177 Ubiquitin-conjugating enzyme E2 binding.
FT REGION 189 195 Free ubiquitin binding.
FT REGION 206 213 Ubiquitin-conjugating enzyme E2 binding.
FT REGION 214 221 Free ubiquitin binding.
FT REGION 245 251 Free ubiquitin binding.
FT ACT_SITE 88 88
FT ACT_SITE 91 91 Nucleophile.
FT ACT_SITE 265 265
FT BINDING 221 221 Free ubiquitin.
FT BINDING 235 235 Free ubiquitin.
FT BINDING 237 237 Free ubiquitin.
FT BINDING 261 261 Free ubiquitin.
FT BINDING 266 266 Free ubiquitin.
FT SITE 23 23 Required for proximal ubiquitin-binding.
FT MOD_RES 2 2 N-acetylalanine.
FT MOD_RES 16 16 Phosphoserine.
FT VAR_SEQ 1 112 MAAEEPQQQKQEPLGSDSEGVNCLAYDEAIMAQQDRIQQEI
FT AVQNPLVSERLELSVLYKEYAEDDNIYQQKIKDLHKKYSYI
FT RKTRPDGNCFYRAFGFSHLEALLDDSKELQ -> MMKPSWL
FT SRTEFSKRLLCRTLWCQSGWSSRSYTRSMLKMTTSINRRSR
FT TSTKSTRTSARPGLTATVSIGLSDSPTWRHCWMTARSCSGE
FT KGGHWAPRQVGVYLLPGRVGCVSSRVSPSFPGDGLDSGLAR
FT RGSAVSALASGLVEEPMLGPPFHPTP (in isoform
FT 2).
FT /FTId=VSP_009464.
FT MUTAGEN 23 23 C->A: Abolishes only ubiquitin-
FT vinylsulfone adduct formation.
FT MUTAGEN 33 33 Q->R: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 37 37 I->T: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 39 39 Q->L: Does not affect activity in DNA
FT repair and ability to inhibit
FT UBE2N/UBC13.
FT MUTAGEN 87 87 P->G: Slightly improves ability to cleave
FT 'K63'-linked ubiquitin.
FT MUTAGEN 88 88 D->E: Abolishes hydrolase activity in
FT vitro. Abolishes ability to inhibit
FT RNF168; when associated with S-91 and A-
FT 265.
FT MUTAGEN 91 91 C->A: Prevents RNF128 autoubiquitination,
FT and stabilizes RNF128 in vivo. Abolishes
FT both ubiquitin-binding and adduct
FT formation with ubiquitin-vinylsulfone.
FT MUTAGEN 91 91 C->S: Abolishes hydrolase activity in
FT vitro. Does not affect ability to inhibit
FT RNF168. Abolishes ability to inhibit
FT RNF168; when associated with A-88 and A-
FT 265.
FT MUTAGEN 116 116 A->T: Does not affect ability to inhibit
FT UBE2N/UBC13.
FT MUTAGEN 134 134 T->R: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 137 137 D->G: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 176 176 R->L: No effect on RNF128.
FT MUTAGEN 190 190 F->S: Fails to inhibit ubiquitin
FT conjugation by UBE2N/UBC13.
FT MUTAGEN 212 212 C->A: No effect on RNF128.
FT MUTAGEN 261 261 Y->H: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 263 263 P->L: Fails to inhibit ubiquitin
FT conjugation by UBE2N/UBC13.
FT MUTAGEN 265 265 H->A: Abolishes ability to inhibit
FT RNF168; when associated with A-88 and S-
FT 91.
FT MUTAGEN 265 265 H->R: Abolishes hydrolase activity in
FT vitro.
FT CONFLICT 61 61 Y -> C (in Ref. 6; AAH10368).
FT CONFLICT 151 151 K -> R (in Ref. 4; BAA90956).
FT HELIX 24 44
FT STRAND 47 53
FT HELIX 54 60
FT HELIX 66 75
FT TURN 76 78
FT STRAND 81 83
FT STRAND 87 89
FT HELIX 91 104
FT HELIX 108 127
FT HELIX 132 150
FT HELIX 155 162
FT HELIX 165 185
FT HELIX 187 190
FT HELIX 191 193
FT STRAND 194 197
FT HELIX 200 207
FT HELIX 217 227
FT STRAND 231 235
FT STRAND 240 242
FT STRAND 245 250
FT STRAND 256 262
FT STRAND 265 270
SQ SEQUENCE 271 AA; 31284 MW; 63188EE1DC5FD66F CRC64;
MAAEEPQQQK QEPLGSDSEG VNCLAYDEAI MAQQDRIQQE IAVQNPLVSE RLELSVLYKE
YAEDDNIYQQ KIKDLHKKYS YIRKTRPDGN CFYRAFGFSH LEALLDDSKE LQRFKAVSAK
SKEDLVSQGF TEFTIEDFHN TFMDLIEQVE KQTSVADLLA SFNDQSTSDY LVVYLRLLTS
GYLQRESKFF EHFIEGGRTV KEFCQQEVEP MCKESDHIHI IALAQALSVS IQVEYMDRGE
GGTTNPHIFP EGSEPKVYLL YRPGHYDILY K
//
ID OTUB1_HUMAN Reviewed; 271 AA.
AC Q96FW1; Q32Q78; Q96II3; Q9NXQ4; Q9P0B8;
DT 01-MAR-2004, integrated into UniProtKB/Swiss-Prot.
read moreDT 01-MAR-2004, sequence version 2.
DT 22-JAN-2014, entry version 115.
DE RecName: Full=Ubiquitin thioesterase OTUB1;
DE EC=3.4.19.12;
DE AltName: Full=Deubiquitinating enzyme OTUB1;
DE AltName: Full=OTU domain-containing ubiquitin aldehyde-binding protein 1;
DE AltName: Full=Otubain-1;
DE Short=hOTU1;
DE AltName: Full=Ubiquitin-specific-processing protease OTUB1;
GN Name=OTUB1; Synonyms=OTB1, OTU1; ORFNames=HSPC263;
OS Homo sapiens (Human).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;
OC Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini;
OC Catarrhini; Hominidae; Homo.
OX NCBI_TaxID=9606;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORM 1), IDENTIFICATION BY MASS
RP SPECTROMETRY, FUNCTION, TISSUE SPECIFICITY, AND MUTAGENESIS OF ASP-88;
RP CYS-91 AND HIS-265.
RC TISSUE=Cervix carcinoma;
RX PubMed=12704427; DOI=10.1038/sj.embor.embor824;
RA Balakirev M.Y., Tcherniuk S.O., Jaquinod M., Chroboczek J.;
RT "Otubains: a new family of cysteine proteases in the ubiquitin
RT pathway.";
RL EMBO Rep. 4:517-522(2003).
RN [2]
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS 1 AND 2), ALTERNATIVE SPLICING,
RP FUNCTION, TISSUE SPECIFICITY, INTERACTION WITH RNF128 AND USP8, AND
RP MUTAGENESIS OF CYS-91; ARG-176 AND CYS-212.
RX PubMed=14661020; DOI=10.1038/ni1017;
RA Soares L., Seroogy C., Skrenta H., Anandasabapathy N., Lovelace P.,
RA Chung C.D., Engleman E., Fathman C.G.;
RT "Two isoforms of otubain 1 regulate T cell anergy via GRAIL.";
RL Nat. Immunol. 5:45-54(2004).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM 1).
RC TISSUE=Umbilical cord blood;
RX PubMed=11042152; DOI=10.1101/gr.140200;
RA Zhang Q.-H., Ye M., Wu X.-Y., Ren S.-X., Zhao M., Zhao C.-J., Fu G.,
RA Shen Y., Fan H.-Y., Lu G., Zhong M., Xu X.-R., Han Z.-G., Zhang J.-W.,
RA Tao J., Huang Q.-H., Zhou J., Hu G.-X., Gu J., Chen S.-J., Chen Z.;
RT "Cloning and functional analysis of cDNAs with open reading frames for
RT 300 previously undefined genes expressed in CD34+ hematopoietic
RT stem/progenitor cells.";
RL Genome Res. 10:1546-1560(2000).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM 1).
RC TISSUE=Colon;
RX PubMed=14702039; DOI=10.1038/ng1285;
RA Ota T., Suzuki Y., Nishikawa T., Otsuki T., Sugiyama T., Irie R.,
RA Wakamatsu A., Hayashi K., Sato H., Nagai K., Kimura K., Makita H.,
RA Sekine M., Obayashi M., Nishi T., Shibahara T., Tanaka T., Ishii S.,
RA Yamamoto J., Saito K., Kawai Y., Isono Y., Nakamura Y., Nagahari K.,
RA Murakami K., Yasuda T., Iwayanagi T., Wagatsuma M., Shiratori A.,
RA Sudo H., Hosoiri T., Kaku Y., Kodaira H., Kondo H., Sugawara M.,
RA Takahashi M., Kanda K., Yokoi T., Furuya T., Kikkawa E., Omura Y.,
RA Abe K., Kamihara K., Katsuta N., Sato K., Tanikawa M., Yamazaki M.,
RA Ninomiya K., Ishibashi T., Yamashita H., Murakawa K., Fujimori K.,
RA Tanai H., Kimata M., Watanabe M., Hiraoka S., Chiba Y., Ishida S.,
RA Ono Y., Takiguchi S., Watanabe S., Yosida M., Hotuta T., Kusano J.,
RA Kanehori K., Takahashi-Fujii A., Hara H., Tanase T.-O., Nomura Y.,
RA Togiya S., Komai F., Hara R., Takeuchi K., Arita M., Imose N.,
RA Musashino K., Yuuki H., Oshima A., Sasaki N., Aotsuka S.,
RA Yoshikawa Y., Matsunawa H., Ichihara T., Shiohata N., Sano S.,
RA Moriya S., Momiyama H., Satoh N., Takami S., Terashima Y., Suzuki O.,
RA Nakagawa S., Senoh A., Mizoguchi H., Goto Y., Shimizu F., Wakebe H.,
RA Hishigaki H., Watanabe T., Sugiyama A., Takemoto M., Kawakami B.,
RA Yamazaki M., Watanabe K., Kumagai A., Itakura S., Fukuzumi Y.,
RA Fujimori Y., Komiyama M., Tashiro H., Tanigami A., Fujiwara T.,
RA Ono T., Yamada K., Fujii Y., Ozaki K., Hirao M., Ohmori Y.,
RA Kawabata A., Hikiji T., Kobatake N., Inagaki H., Ikema Y., Okamoto S.,
RA Okitani R., Kawakami T., Noguchi S., Itoh T., Shigeta K., Senba T.,
RA Matsumura K., Nakajima Y., Mizuno T., Morinaga M., Sasaki M.,
RA Togashi T., Oyama M., Hata H., Watanabe M., Komatsu T.,
RA Mizushima-Sugano J., Satoh T., Shirai Y., Takahashi Y., Nakagawa K.,
RA Okumura K., Nagase T., Nomura N., Kikuchi H., Masuho Y., Yamashita R.,
RA Nakai K., Yada T., Nakamura Y., Ohara O., Isogai T., Sugano S.;
RT "Complete sequencing and characterization of 21,243 full-length human
RT cDNAs.";
RL Nat. Genet. 36:40-45(2004).
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RX PubMed=16554811; DOI=10.1038/nature04632;
RA Taylor T.D., Noguchi H., Totoki Y., Toyoda A., Kuroki Y., Dewar K.,
RA Lloyd C., Itoh T., Takeda T., Kim D.-W., She X., Barlow K.F.,
RA Bloom T., Bruford E., Chang J.L., Cuomo C.A., Eichler E.,
RA FitzGerald M.G., Jaffe D.B., LaButti K., Nicol R., Park H.-S.,
RA Seaman C., Sougnez C., Yang X., Zimmer A.R., Zody M.C., Birren B.W.,
RA Nusbaum C., Fujiyama A., Hattori M., Rogers J., Lander E.S.,
RA Sakaki Y.;
RT "Human chromosome 11 DNA sequence and analysis including novel gene
RT identification.";
RL Nature 440:497-500(2006).
RN [6]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] (ISOFORM 1).
RC TISSUE=Colon, and Skin;
RX PubMed=15489334; DOI=10.1101/gr.2596504;
RG The MGC Project Team;
RT "The status, quality, and expansion of the NIH full-length cDNA
RT project: the Mammalian Gene Collection (MGC).";
RL Genome Res. 14:2121-2127(2004).
RN [7]
RP PARTIAL PROTEIN SEQUENCE, AND FUNCTION.
RX PubMed=12401499; DOI=10.1016/S1074-5521(02)00248-X;
RA Borodovsky A., Ovaa H., Kolli N., Gan-Erdene T., Wilkinson K.D.,
RA Ploegh H.L., Kessler B.M.;
RT "Chemistry-based functional proteomics reveals novel members of the
RT deubiquitinating enzyme family.";
RL Chem. Biol. 9:1149-1159(2002).
RN [8]
RP PHOSPHORYLATION [LARGE SCALE ANALYSIS] AT SER-16, AND MASS
RP SPECTROMETRY.
RC TISSUE=Cervix carcinoma;
RX PubMed=18669648; DOI=10.1073/pnas.0805139105;
RA Dephoure N., Zhou C., Villen J., Beausoleil S.A., Bakalarski C.E.,
RA Elledge S.J., Gygi S.P.;
RT "A quantitative atlas of mitotic phosphorylation.";
RL Proc. Natl. Acad. Sci. U.S.A. 105:10762-10767(2008).
RN [9]
RP ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, MASS SPECTROMETRY, AND
RP CLEAVAGE OF INITIATOR METHIONINE.
RX PubMed=19413330; DOI=10.1021/ac9004309;
RA Gauci S., Helbig A.O., Slijper M., Krijgsveld J., Heck A.J.,
RA Mohammed S.;
RT "Lys-N and trypsin cover complementary parts of the phosphoproteome in
RT a refined SCX-based approach.";
RL Anal. Chem. 81:4493-4501(2009).
RN [10]
RP FUNCTION, INTERACTION WITH ESR1, AND MUTAGENESIS OF CYS-91.
RX PubMed=19383985; DOI=10.1074/jbc.M109.007484;
RA Stanisic V., Malovannaya A., Qin J., Lonard D.M., O'Malley B.W.;
RT "OTU Domain-containing ubiquitin aldehyde-binding protein 1 (OTUB1)
RT deubiquitinates estrogen receptor (ER) alpha and affects ERalpha
RT transcriptional activity.";
RL J. Biol. Chem. 284:16135-16145(2009).
RN [11]
RP FUNCTION, UBIQUITIN-BINDING, DOMAIN, AND MUTAGENESIS OF CYS-23; CYS-91
RP AND CYS-212.
RX PubMed=19211026; DOI=10.1016/j.jmb.2008.12.085;
RA Wang T., Yin L., Cooper E.M., Lai M.-Y., Dickey S., Pickart C.M.,
RA Fushman D., Wilkinson K.D., Cohen R.E., Wolberger C.;
RT "Evidence for bidentate substrate binding as the basis for the K48
RT linkage specificity of otubain 1.";
RL J. Mol. Biol. 386:1011-1023(2009).
RN [12]
RP FUNCTION IN INHIBITION OF RNF168, INTERACTION WITH UBE2N/UBC13, AND
RP MUTAGENESIS OF ASP-88; CYS-91 AND HIS-265.
RX PubMed=20725033; DOI=10.1038/nature09297;
RA Nakada S., Tai I., Panier S., Al-Hakim A., Iemura S., Juang Y.C.,
RA O'Donnell L., Kumakubo A., Munro M., Sicheri F., Gingras A.C.,
RA Natsume T., Suda T., Durocher D.;
RT "Non-canonical inhibition of DNA damage-dependent ubiquitination by
RT OTUB1.";
RL Nature 466:941-946(2010).
RN [13]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RX PubMed=21269460; DOI=10.1186/1752-0509-5-17;
RA Burkard T.R., Planyavsky M., Kaupe I., Breitwieser F.P.,
RA Buerckstuemmer T., Bennett K.L., Superti-Furga G., Colinge J.;
RT "Initial characterization of the human central proteome.";
RL BMC Syst. Biol. 5:17-17(2011).
RN [14]
RP ACETYLATION [LARGE SCALE ANALYSIS] AT ALA-2, AND MASS SPECTROMETRY.
RX PubMed=22814378; DOI=10.1073/pnas.1210303109;
RA Van Damme P., Lasa M., Polevoda B., Gazquez C., Elosegui-Artola A.,
RA Kim D.S., De Juan-Pardo E., Demeyer K., Hole K., Larrea E.,
RA Timmerman E., Prieto J., Arnesen T., Sherman F., Gevaert K.,
RA Aldabe R.;
RT "N-terminal acetylome analyses and functional insights of the N-
RT terminal acetyltransferase NatB.";
RL Proc. Natl. Acad. Sci. U.S.A. 109:12449-12454(2012).
RN [15]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=23827681; DOI=10.1016/j.cell.2013.05.046;
RA Mevissen T.E., Hospenthal M.K., Geurink P.P., Elliott P.R., Akutsu M.,
RA Arnaudo N., Ekkebus R., Kulathu Y., Wauer T., El Oualid F.,
RA Freund S.M., Ovaa H., Komander D.;
RT "OTU deubiquitinases reveal mechanisms of linkage specificity and
RT enable ubiquitin chain restriction analysis.";
RL Cell 154:169-184(2013).
RN [16]
RP X-RAY CRYSTALLOGRAPHY (1.69 ANGSTROMS) OF 40-271, FUNCTION,
RP INTERACTION WITH FUS AND GNB2L1, AND MUTAGENESIS OF PRO-87 AND CYS-91.
RX PubMed=18954305; DOI=10.1042/BJ20081318;
RA Edelmann M.J., Iphoefer A., Akutsu M., Altun M., di Gleria K.,
RA Kramer H.B., Fiebiger E., Dhe-Paganon S., Kessler B.M.;
RT "Structural basis and specificity of human otubain 1-mediated
RT deubiquitination.";
RL Biochem. J. 418:379-390(2009).
RN [17]
RP X-RAY CRYSTALLOGRAPHY (3.3 ANGSTROMS) OF 25-271 IN COMPLEX WITH UBE2V2
RP AND UBE2N, FREE UBIQUITIN-BINDING, ENZYME REGULATION, AND MUTAGENESIS
RP OF GLN-33; ILE-37; GLN-39; CYS-91; ALA-116; THR-134; ASP-137; PHE-190;
RP TYR-261 AND PRO-263.
RX PubMed=22325355; DOI=10.1016/j.molcel.2012.01.011;
RA Juang Y.C., Landry M.C., Sanches M., Vittal V., Leung C.C.,
RA Ceccarelli D.F., Mateo A.R., Pruneda J.N., Mao D.Y., Szilard R.K.,
RA Orlicky S., Munro M., Brzovic P.S., Klevit R.E., Sicheri F.,
RA Durocher D.;
RT "OTUB1 co-opts Lys48-linked ubiquitin recognition to suppress E2
RT enzyme function.";
RL Mol. Cell 45:384-397(2012).
RN [18]
RP X-RAY CRYSTALLOGRAPHY (3.11 ANGSTROMS) OF 1-45 IN COMPLEX WITH UBE2N
RP AND UBIQUITIN, ENZYME REGULATION, AND FREE UBIQUITIN-BINDING.
RX PubMed=22367539; DOI=10.1038/nature10911;
RA Wiener R., Zhang X., Wang T., Wolberger C.;
RT "The mechanism of OTUB1-mediated inhibition of ubiquitination.";
RL Nature 483:618-622(2012).
CC -!- FUNCTION: Hydrolase that can specifically remove 'Lys-48'-linked
CC conjugated ubiquitin from proteins and plays an important
CC regulatory role at the level of protein turnover by preventing
CC degradation. Regulator of T-cell anergy, a phenomenon that occurs
CC when T-cells are rendered unresponsive to antigen rechallenge and
CC no longer respond to their cognate antigen. Acts via its
CC interaction with RNF128/GRAIL, a crucial inductor of CD4 T-cell
CC anergy. Isoform 1 destabilizes RNF128, leading to prevent anergy.
CC In contrast, isoform 2 stabilizes RNF128 and promotes anergy.
CC Surprisingly, it regulates RNF128-mediated ubiquitination, but
CC does not deubiquitinate polyubiquitinated RNF128. Deubiquitinates
CC estrogen receptor alpha (ESR1). Mediates deubiquitination of 'Lys-
CC 48'-linked polyubiquitin chains, but not 'Lys-63'-linked
CC polyubiquitin chains. Not able to cleave di-ubiquitin. Also
CC capable of removing NEDD8 from NEDD8 conjugates, but with a much
CC lower preference compared to 'Lys-48'-linked ubiquitin.
CC -!- FUNCTION: Plays a key non-catalytic role in DNA repair regulation
CC by inhibiting activity of RNF168, an E3 ubiquitin-protein ligase
CC that promotes accumulation of 'Lys-63'-linked histone H2A and H2AX
CC at DNA damage sites. Inhibits RNF168 independently of ubiquitin
CC thioesterase activity by binding and inhibiting UBE2N/UBC13, the
CC E2 partner of RNF168, thereby limiting spreading of 'Lys-63'-
CC linked histone H2A and H2AX marks. Inhibition occurs by binding to
CC free ubiquitin: free ubiquitin acts as an allosteric regulator
CC that increases affinity for UBE2N/UBC13 and disrupts interaction
CC with UBE2V1. The OTUB1-UBE2N/UBC13-free ubiquitin complex adopts a
CC configuration that mimics a cleaved 'Lys48'-linked di-ubiquitin
CC chain.
CC -!- CATALYTIC ACTIVITY: Thiol-dependent hydrolysis of ester,
CC thioester, amide, peptide and isopeptide bonds formed by the C-
CC terminal Gly of ubiquitin (a 76-residue protein attached to
CC proteins as an intracellular targeting signal).
CC -!- ENZYME REGULATION: By free ubiquitin: binding of free ubiquitin
CC triggers conformational changes in the OTU domain and formation of
CC a ubiquitin-binding helix in the N-terminus, promoting binding of
CC the conjugated donor ubiquitin in UBE2N/UBC13 to OTUB1.
CC -!- SUBUNIT: Isoform 1 and isoform 2 interact with RNF128. Isoform 1
CC forms a ternary complex with RNF128 and USP8. Isoform 1 interacts
CC with the C-terminal UCH catalytic domain of USP8. Isoform 2 does
CC not associate with USP8. Interacts with FUS, ESR1 and
CC GNB2L1/RACK1. Interacts with UBE2N/UBC13.
CC -!- INTERACTION:
CC Q13490:BIRC2; NbExp=3; IntAct=EBI-1058491, EBI-514538;
CC Q00987:MDM2; NbExp=5; IntAct=EBI-1058491, EBI-389668;
CC P04637:TP53; NbExp=8; IntAct=EBI-1058491, EBI-366083;
CC P51668:UBE2D1; NbExp=4; IntAct=EBI-1058491, EBI-743540;
CC P62837:UBE2D2; NbExp=5; IntAct=EBI-1058491, EBI-347677;
CC P61077:UBE2D3; NbExp=4; IntAct=EBI-1058491, EBI-348268;
CC Q56921:ypkA (xeno); NbExp=5; IntAct=EBI-1058491, EBI-8022937;
CC Q9RI12:ypkA (xeno); NbExp=3; IntAct=EBI-1058491, EBI-2849107;
CC -!- SUBCELLULAR LOCATION: Cytoplasm (By similarity).
CC -!- ALTERNATIVE PRODUCTS:
CC Event=Alternative splicing; Named isoforms=2;
CC Name=1; Synonyms=Otubain-1;
CC IsoId=Q96FW1-1; Sequence=Displayed;
CC Name=2; Synonyms=ARF-1;
CC IsoId=Q96FW1-2; Sequence=VSP_009464;
CC Note=Lacks the catalytic sites for protease activity;
CC -!- TISSUE SPECIFICITY: Isoform 1 is ubiquitous. Isoform 2 is
CC expressed only in lymphoid tissues such as tonsils, lymph nodes
CC and spleen, as well as peripheral blood mononuclear cells.
CC -!- DOMAIN: In addition to ubiquitin-binding at the Cys-91 active
CC site, a proximal ubiquitin-binding site is also present at Cys-23
CC Occupancy of the active site is needed to enable tight binding to
CC the second site. Distinct binding sites for the ubiquitins may
CC allow to discriminate among different isopeptide linkages (i.e.
CC 'Lys-48'-, 'Lys-63'-linked polyubiquitin) in polyubiquitin
CC substrates and achieve linkage-specific deubiquitination.
CC -!- MISCELLANEOUS: In the structure described by PubMed:18954305, the
CC His-265 active site of the catalytic triad is located too far to
CC interact directly with the active site Cys-91. A possible
CC explanation is that OTUB1 is in inactive conformation in absence
CC of ubiquitin and a conformation change may move His-265 in the
CC proximity of Cys-91 in presence of ubiquitin substrate.
CC -!- SIMILARITY: Belongs to the peptidase C65 family.
CC -!- SIMILARITY: Contains 1 OTU domain.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAF28941.1; Type=Erroneous initiation;
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DR EMBL; AY177200; AAO27702.1; -; mRNA.
DR EMBL; AF161381; AAF28941.1; ALT_INIT; mRNA.
DR EMBL; AK000120; BAA90956.1; -; mRNA.
DR EMBL; AP000721; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR EMBL; BC007519; AAH07519.1; -; mRNA.
DR EMBL; BC010368; AAH10368.1; -; mRNA.
DR EMBL; BC107701; AAI07702.1; -; mRNA.
DR RefSeq; NP_060140.2; NM_017670.2.
DR UniGene; Hs.473788; -.
DR PDB; 2ZFY; X-ray; 1.69 A; A=40-271.
DR PDB; 3VON; X-ray; 3.15 A; A/H/O/V/c/j=45-271.
DR PDB; 4DDG; X-ray; 3.30 A; A/B/C/J/K/L=25-271.
DR PDB; 4DDI; X-ray; 3.80 A; A/B/C=25-271.
DR PDB; 4DHZ; X-ray; 3.11 A; A=1-45.
DR PDB; 4I6L; X-ray; 2.49 A; A=45-271.
DR PDB; 4LDT; X-ray; 1.90 A; A=1-45.
DR PDBsum; 2ZFY; -.
DR PDBsum; 3VON; -.
DR PDBsum; 4DDG; -.
DR PDBsum; 4DDI; -.
DR PDBsum; 4DHZ; -.
DR PDBsum; 4I6L; -.
DR PDBsum; 4LDT; -.
DR ProteinModelPortal; Q96FW1; -.
DR SMR; Q96FW1; 25-271.
DR DIP; DIP-41158N; -.
DR IntAct; Q96FW1; 26.
DR MINT; MINT-5001867; -.
DR STRING; 9606.ENSP00000301453; -.
DR MEROPS; C65.001; -.
DR PhosphoSite; Q96FW1; -.
DR DMDM; 44888286; -.
DR PaxDb; Q96FW1; -.
DR PRIDE; Q96FW1; -.
DR DNASU; 55611; -.
DR Ensembl; ENST00000428192; ENSP00000402551; ENSG00000167770.
DR Ensembl; ENST00000538426; ENSP00000444357; ENSG00000167770.
DR GeneID; 55611; -.
DR KEGG; hsa:55611; -.
DR UCSC; uc001nyf.1; human.
DR CTD; 55611; -.
DR GeneCards; GC11P063753; -.
DR HGNC; HGNC:23077; OTUB1.
DR HPA; HPA039176; -.
DR MIM; 608337; gene.
DR neXtProt; NX_Q96FW1; -.
DR PharmGKB; PA134988141; -.
DR eggNOG; NOG267426; -.
DR HOVERGEN; HBG053383; -.
DR InParanoid; Q96FW1; -.
DR KO; K09602; -.
DR PhylomeDB; Q96FW1; -.
DR ChiTaRS; OTUB1; human.
DR EvolutionaryTrace; Q96FW1; -.
DR GeneWiki; OTUB1; -.
DR GenomeRNAi; 55611; -.
DR NextBio; 60179; -.
DR PRO; PR:Q96FW1; -.
DR ArrayExpress; Q96FW1; -.
DR Bgee; Q96FW1; -.
DR CleanEx; HS_OTUB1; -.
DR Genevestigator; Q96FW1; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0004843; F:deubiquitinase activity; IDA:UniProtKB.
DR GO; GO:0019784; F:NEDD8-specific protease activity; IDA:UniProtKB.
DR GO; GO:0008242; F:omega peptidase activity; IEA:InterPro.
DR GO; GO:0043130; F:ubiquitin binding; IDA:UniProtKB.
DR GO; GO:0006974; P:cellular response to DNA damage stimulus; IDA:UniProtKB.
DR GO; GO:0006281; P:DNA repair; IEA:UniProtKB-KW.
DR GO; GO:2000780; P:negative regulation of double-strand break repair; IMP:UniProtKB.
DR GO; GO:1901315; P:negative regulation of histone H2A K63-linked ubiquitination; IDA:UniProtKB.
DR GO; GO:0071108; P:protein K48-linked deubiquitination; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR InterPro; IPR003323; OTU.
DR InterPro; IPR019400; Peptidase_C65_otubain.
DR InterPro; IPR016615; Ubiquitin_thioesterase_Otubain.
DR Pfam; PF10275; Peptidase_C65; 1.
DR PIRSF; PIRSF013503; Ubiquitin_thioesterase_Otubain; 1.
DR PROSITE; PS50802; OTU; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Acetylation; Adaptive immunity; Alternative splicing;
KW Complete proteome; Cytoplasm; Direct protein sequencing; DNA damage;
KW DNA repair; Hydrolase; Immunity; Phosphoprotein; Protease;
KW Reference proteome; Thiol protease; Ubl conjugation pathway.
FT INIT_MET 1 1 Removed.
FT CHAIN 2 271 Ubiquitin thioesterase OTUB1.
FT /FTId=PRO_0000221008.
FT DOMAIN 80 271 OTU.
FT REGION 130 138 Ubiquitin-conjugating enzyme E2 binding.
FT REGION 169 177 Ubiquitin-conjugating enzyme E2 binding.
FT REGION 189 195 Free ubiquitin binding.
FT REGION 206 213 Ubiquitin-conjugating enzyme E2 binding.
FT REGION 214 221 Free ubiquitin binding.
FT REGION 245 251 Free ubiquitin binding.
FT ACT_SITE 88 88
FT ACT_SITE 91 91 Nucleophile.
FT ACT_SITE 265 265
FT BINDING 221 221 Free ubiquitin.
FT BINDING 235 235 Free ubiquitin.
FT BINDING 237 237 Free ubiquitin.
FT BINDING 261 261 Free ubiquitin.
FT BINDING 266 266 Free ubiquitin.
FT SITE 23 23 Required for proximal ubiquitin-binding.
FT MOD_RES 2 2 N-acetylalanine.
FT MOD_RES 16 16 Phosphoserine.
FT VAR_SEQ 1 112 MAAEEPQQQKQEPLGSDSEGVNCLAYDEAIMAQQDRIQQEI
FT AVQNPLVSERLELSVLYKEYAEDDNIYQQKIKDLHKKYSYI
FT RKTRPDGNCFYRAFGFSHLEALLDDSKELQ -> MMKPSWL
FT SRTEFSKRLLCRTLWCQSGWSSRSYTRSMLKMTTSINRRSR
FT TSTKSTRTSARPGLTATVSIGLSDSPTWRHCWMTARSCSGE
FT KGGHWAPRQVGVYLLPGRVGCVSSRVSPSFPGDGLDSGLAR
FT RGSAVSALASGLVEEPMLGPPFHPTP (in isoform
FT 2).
FT /FTId=VSP_009464.
FT MUTAGEN 23 23 C->A: Abolishes only ubiquitin-
FT vinylsulfone adduct formation.
FT MUTAGEN 33 33 Q->R: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 37 37 I->T: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 39 39 Q->L: Does not affect activity in DNA
FT repair and ability to inhibit
FT UBE2N/UBC13.
FT MUTAGEN 87 87 P->G: Slightly improves ability to cleave
FT 'K63'-linked ubiquitin.
FT MUTAGEN 88 88 D->E: Abolishes hydrolase activity in
FT vitro. Abolishes ability to inhibit
FT RNF168; when associated with S-91 and A-
FT 265.
FT MUTAGEN 91 91 C->A: Prevents RNF128 autoubiquitination,
FT and stabilizes RNF128 in vivo. Abolishes
FT both ubiquitin-binding and adduct
FT formation with ubiquitin-vinylsulfone.
FT MUTAGEN 91 91 C->S: Abolishes hydrolase activity in
FT vitro. Does not affect ability to inhibit
FT RNF168. Abolishes ability to inhibit
FT RNF168; when associated with A-88 and A-
FT 265.
FT MUTAGEN 116 116 A->T: Does not affect ability to inhibit
FT UBE2N/UBC13.
FT MUTAGEN 134 134 T->R: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 137 137 D->G: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 176 176 R->L: No effect on RNF128.
FT MUTAGEN 190 190 F->S: Fails to inhibit ubiquitin
FT conjugation by UBE2N/UBC13.
FT MUTAGEN 212 212 C->A: No effect on RNF128.
FT MUTAGEN 261 261 Y->H: Impairs inhibition of UBE2N/UBC13.
FT MUTAGEN 263 263 P->L: Fails to inhibit ubiquitin
FT conjugation by UBE2N/UBC13.
FT MUTAGEN 265 265 H->A: Abolishes ability to inhibit
FT RNF168; when associated with A-88 and S-
FT 91.
FT MUTAGEN 265 265 H->R: Abolishes hydrolase activity in
FT vitro.
FT CONFLICT 61 61 Y -> C (in Ref. 6; AAH10368).
FT CONFLICT 151 151 K -> R (in Ref. 4; BAA90956).
FT HELIX 24 44
FT STRAND 47 53
FT HELIX 54 60
FT HELIX 66 75
FT TURN 76 78
FT STRAND 81 83
FT STRAND 87 89
FT HELIX 91 104
FT HELIX 108 127
FT HELIX 132 150
FT HELIX 155 162
FT HELIX 165 185
FT HELIX 187 190
FT HELIX 191 193
FT STRAND 194 197
FT HELIX 200 207
FT HELIX 217 227
FT STRAND 231 235
FT STRAND 240 242
FT STRAND 245 250
FT STRAND 256 262
FT STRAND 265 270
SQ SEQUENCE 271 AA; 31284 MW; 63188EE1DC5FD66F CRC64;
MAAEEPQQQK QEPLGSDSEG VNCLAYDEAI MAQQDRIQQE IAVQNPLVSE RLELSVLYKE
YAEDDNIYQQ KIKDLHKKYS YIRKTRPDGN CFYRAFGFSH LEALLDDSKE LQRFKAVSAK
SKEDLVSQGF TEFTIEDFHN TFMDLIEQVE KQTSVADLLA SFNDQSTSDY LVVYLRLLTS
GYLQRESKFF EHFIEGGRTV KEFCQQEVEP MCKESDHIHI IALAQALSVS IQVEYMDRGE
GGTTNPHIFP EGSEPKVYLL YRPGHYDILY K
//
MIM
608337
*RECORD*
*FIELD* NO
608337
*FIELD* TI
*608337 OTU DOMAIN-CONTAINING UBIQUITIN ALDEHYDE-BINDING PROTEIN 1; OTUB1
;;OTU DOMAIN-CONTAINING UBAL-BINDING PROTEIN 1;;
read moreOTUBAIN 1; OTU1; OTB1
*FIELD* TX
DESCRIPTION
Otubains, such as OTB1, are deubiquitylating cysteine proteases (DUBs;
see 602519) that belong to the ovarian tumor (OTU) protein superfamily.
Like other DUBs, otubains cleave proteins precisely at the ubiquitin
(UB; see 191339)-protein bond (Balakirev et al., 2003).
CLONING
By affinity purification of DUBs from HeLa cells using UB aldehyde
(UBAL), followed by SDS-PAGE and mass spectrophotometric analysis,
Balakirev et al. (2003) obtained cDNAs encoding OTB1 and OTB2 (608338).
The 271-amino acid, 31-kD OTB1 protein contains UB-interaction motifs
(UIMs) and UB-associated (UBA) domains, as well as putative nuclear
localization signals and a consensus LxxLL motif. RT-PCR and Western
blot analyses revealed wide expression of OTB1, with 2 isoforms in
leukocytes.
By PCR and immunoblot analysis, Soares et al. (2004) identified an
815-bp OTB1 isoform, which was expressed ubiquitously, and a 950-bp
isoform, which was expressed only in lymphoid tissue. The 815-bp isoform
encodes the 31-kD protein reported by Balakirev et al. (2003), whereas
the 950-bp isoform encodes a 35-kD, 315-amino acid protein, OTB1ARF1
(OTB1 alternative reading frame-1), that lacks the asp and cys residues
in the OTU cysteine proteinase catalytic core and retains only the
C-terminal flanking region of the OTU domain. The OTB1ARF1 transcript is
produced from the same transcription start site as OTB1 but uses an AUG
codon located in exon 2 and in a different reading frame from that of
OTB1. In addition, there is no splicing between exons 4 and 5 in
OTB1ARF1, which causes a frameshift back to the same reading frame as
OTB1.
GENE FUNCTION
By yeast 2-hybrid analysis, Soares et al. (2004) showed that GRAIL
(RNF128; 300439) binds to OTB1. Although OTB1 was found to have DUB
activity, it failed to deubiquitinate polyubiquitinated GRAIL.
Immunoprecipitation and coexpression analysis indicated that OTB1, but
not OTB1ARF1, binds to the C-terminal UCH catalytic domain of USP8
(603158) in a trimolecular complex with GRAIL. Soares et al. (2004)
studied cells from T-cell receptor-transgenic mice expressing the
alternative forms of OTB1. OTB1-expressing cells contained negligible
amounts of Grail, proliferated well, and produced large amounts of
interleukin-2 (IL2; 147680) after antigenic stimulation. In contrast,
cells expressing OTB1ARF1 contained large amounts of Grail and were
functionally anergic, proliferating poorly and not producing Il2. Soares
et al. (2004) concluded that the balance of OTB1 and OTB1ARF1 in T cells
is an important factor that regulates GRAIL stability and the outcome of
immune responses in secondary lymphoid organs.
Nakada et al. (2010) reported that OTUB1, a deubiquitinating enzyme, is
an inhibitor of double stranded break induced chromatin ubiquitination.
Surprisingly, they found that OTUB1 suppresses RNF168 (612688)-dependent
polyubiquitination independently of its catalytic activity. OTUB1 does
so by binding to and inhibiting UBC13 (UBE2N; 603679), the cognate E2
enzyme for RNF168. Nakada et al. (2010) suggested that this unusual mode
of regulation is unlikely to be limited to UBC13 because analysis of
OTUB1-associated proteins revealed that OTUB1 binds to E2s of the UBE2D
and UBE2E subfamilies. Finally, OTUB1 depletion mitigates the
double-stranded break repair defect associated with defective ATM
(607585) signaling, indicating that pharmacologic targeting of the
OTUB1-UBC13 interaction might enhance the DNA damage response.
Using immunoblot analysis, Lin et al. (2009) demonstrated expression of
GRAIL in naive mouse and human CD4 (186940) T lymphocytes. They found
that, upon CD4 T-cell activation, OTUB1 was expressed and GRAIL was
degraded, allowing proliferation to proceed. Preventing OTUB1 expression
with CTLA4 (123890)-Ig, anti-IL2, or rapamycin, all of which target MTOR
(FRAP1; 601231), resulted in maintenance of GRAIL and inhibition of CD4
T-cell proliferation. Lin et al. (2009) concluded that a pathway
sequentially links CD28 (186760) costimulation, IL2 expression, IL2R
(IL2RA; 147730) signaling, and MTOR activation as important requirements
for naive CD4 T-cell proliferation through the regulation of OTUB1 and
GRAIL expression.
GENE STRUCTURE
Soares et al. (2004) determined that the OTB1 gene contains 7 exons.
BIOCHEMICAL FEATURES
- Crystal Structure
Wiener et al. (2012) described structural and biochemical studies
elucidating how OTUB1 inhibits UBC13 and other E2 enzymes. They
unexpectedly found that OTUB1 binding to UBC13-Ub (the ubiquitin
thiolester) is allosterically regulated by free ubiquitin, which binds
to a second site in OTUB1 and increases its affinity for UBC13-Ub, while
at the same time disrupting interactions with UEV1A (602995) in a manner
that depends on the OTUB1 N terminus. Crystal structures of an
OTUB1-UBC13 complex and of OTUB1 bound to ubiquitin aldehyde and a
chemical UBC13-Ub conjugate showed that binding of free ubiquitin to
OTUB1 triggers conformational changes in the OTU domain and formation of
a ubiquitin-binding helix in the N terminus, thus promoting binding of
the conjugated donor ubiquitin in UBC13-Ub to OTUB1. The donor ubiquitin
thus cannot interact with the E2 enzyme, which has been shown to be
important for ubiquitin transfer. The N-terminal helix of OTUB1 is
positioned to interfere with UEV1A binding to UBC13, as well as with
attack on the thiolester by an acceptor ubiquitin, thereby inhibiting
K63Ub synthesis. OTUB1 binding also occludes the RING E3 binding site on
UBC13, thus providing a further component of inhibition. Wiener et al.
(2012) concluded that the general features of the inhibition mechanism
explained how OTUB1 inhibits other E2 enzymes in a noncatalytic manner.
MAPPING
By genomic sequence analysis, Balakirev et al. (2003) mapped the OTUB1
gene to chromosome 11q13.1.
*FIELD* RF
1. Balakirev, M. Y.; Tcherniuk, S. O.; Jaquinod, M.; Chroboczek, J.
: Otubains: a new family of cysteine proteases in the ubiquitin pathway. EMBO
Reports 4: 517-522, 2003.
2. Lin, J. T.; Lineberry, N. B.; Kattah, M. G.; Su, L. L.; Utz, P.
J.; Fathman, C. G.; Wu, L.: Naive CD4 T cell proliferation is controlled
by mammalian target of rapamycin regulation of GRAIL expression. J.
Immun. 182: 5919-5928, 2009.
3. Nakada, S.; Tai, I.; Panier, S.; Al-Hakim, A.; Iemura, S.; Juang,
Y.-C.; O'Donnell, L.; Kumakubo, A.; Munro, M.; Sicheri, F.; Gingras,
A.-C.; Natsume, T.; Suda, T.; Durocher, D.: Non-canonical inhibition
of DNA damage-dependent ubiquitination by OTUB1. Nature 466: 941-946,
2010.
4. Soares, L.; Seroogy, C.; Skrenta, H.; Anandasabapathy, N.; Lovelace,
P.; Chung, C. D.; Engleman, E.; Fathman, C. G.: Two isoforms of otubain
1 regulate T cell anergy via GRAIL. Nature Immun. 5: 45-54, 2004.
5. Wiener, R.; Zhang, X.; Wang, T.; Wolberger, C.: The mechanism
of OTUB1-mediated inhibition of ubiquitination. Nature 483: 618-622,
2012.
*FIELD* CN
Ada Hamosh - updated: 5/15/2012
Paul J. Converse - updated: 2/28/2011
Ada Hamosh - updated: 9/21/2010
*FIELD* CD
Paul J. Converse: 12/11/2003
*FIELD* ED
alopez: 05/16/2012
alopez: 5/16/2012
terry: 5/15/2012
mgross: 3/1/2011
terry: 2/28/2011
alopez: 9/22/2010
terry: 9/21/2010
alopez: 5/6/2008
alopez: 2/18/2004
mgross: 12/11/2003
*RECORD*
*FIELD* NO
608337
*FIELD* TI
*608337 OTU DOMAIN-CONTAINING UBIQUITIN ALDEHYDE-BINDING PROTEIN 1; OTUB1
;;OTU DOMAIN-CONTAINING UBAL-BINDING PROTEIN 1;;
read moreOTUBAIN 1; OTU1; OTB1
*FIELD* TX
DESCRIPTION
Otubains, such as OTB1, are deubiquitylating cysteine proteases (DUBs;
see 602519) that belong to the ovarian tumor (OTU) protein superfamily.
Like other DUBs, otubains cleave proteins precisely at the ubiquitin
(UB; see 191339)-protein bond (Balakirev et al., 2003).
CLONING
By affinity purification of DUBs from HeLa cells using UB aldehyde
(UBAL), followed by SDS-PAGE and mass spectrophotometric analysis,
Balakirev et al. (2003) obtained cDNAs encoding OTB1 and OTB2 (608338).
The 271-amino acid, 31-kD OTB1 protein contains UB-interaction motifs
(UIMs) and UB-associated (UBA) domains, as well as putative nuclear
localization signals and a consensus LxxLL motif. RT-PCR and Western
blot analyses revealed wide expression of OTB1, with 2 isoforms in
leukocytes.
By PCR and immunoblot analysis, Soares et al. (2004) identified an
815-bp OTB1 isoform, which was expressed ubiquitously, and a 950-bp
isoform, which was expressed only in lymphoid tissue. The 815-bp isoform
encodes the 31-kD protein reported by Balakirev et al. (2003), whereas
the 950-bp isoform encodes a 35-kD, 315-amino acid protein, OTB1ARF1
(OTB1 alternative reading frame-1), that lacks the asp and cys residues
in the OTU cysteine proteinase catalytic core and retains only the
C-terminal flanking region of the OTU domain. The OTB1ARF1 transcript is
produced from the same transcription start site as OTB1 but uses an AUG
codon located in exon 2 and in a different reading frame from that of
OTB1. In addition, there is no splicing between exons 4 and 5 in
OTB1ARF1, which causes a frameshift back to the same reading frame as
OTB1.
GENE FUNCTION
By yeast 2-hybrid analysis, Soares et al. (2004) showed that GRAIL
(RNF128; 300439) binds to OTB1. Although OTB1 was found to have DUB
activity, it failed to deubiquitinate polyubiquitinated GRAIL.
Immunoprecipitation and coexpression analysis indicated that OTB1, but
not OTB1ARF1, binds to the C-terminal UCH catalytic domain of USP8
(603158) in a trimolecular complex with GRAIL. Soares et al. (2004)
studied cells from T-cell receptor-transgenic mice expressing the
alternative forms of OTB1. OTB1-expressing cells contained negligible
amounts of Grail, proliferated well, and produced large amounts of
interleukin-2 (IL2; 147680) after antigenic stimulation. In contrast,
cells expressing OTB1ARF1 contained large amounts of Grail and were
functionally anergic, proliferating poorly and not producing Il2. Soares
et al. (2004) concluded that the balance of OTB1 and OTB1ARF1 in T cells
is an important factor that regulates GRAIL stability and the outcome of
immune responses in secondary lymphoid organs.
Nakada et al. (2010) reported that OTUB1, a deubiquitinating enzyme, is
an inhibitor of double stranded break induced chromatin ubiquitination.
Surprisingly, they found that OTUB1 suppresses RNF168 (612688)-dependent
polyubiquitination independently of its catalytic activity. OTUB1 does
so by binding to and inhibiting UBC13 (UBE2N; 603679), the cognate E2
enzyme for RNF168. Nakada et al. (2010) suggested that this unusual mode
of regulation is unlikely to be limited to UBC13 because analysis of
OTUB1-associated proteins revealed that OTUB1 binds to E2s of the UBE2D
and UBE2E subfamilies. Finally, OTUB1 depletion mitigates the
double-stranded break repair defect associated with defective ATM
(607585) signaling, indicating that pharmacologic targeting of the
OTUB1-UBC13 interaction might enhance the DNA damage response.
Using immunoblot analysis, Lin et al. (2009) demonstrated expression of
GRAIL in naive mouse and human CD4 (186940) T lymphocytes. They found
that, upon CD4 T-cell activation, OTUB1 was expressed and GRAIL was
degraded, allowing proliferation to proceed. Preventing OTUB1 expression
with CTLA4 (123890)-Ig, anti-IL2, or rapamycin, all of which target MTOR
(FRAP1; 601231), resulted in maintenance of GRAIL and inhibition of CD4
T-cell proliferation. Lin et al. (2009) concluded that a pathway
sequentially links CD28 (186760) costimulation, IL2 expression, IL2R
(IL2RA; 147730) signaling, and MTOR activation as important requirements
for naive CD4 T-cell proliferation through the regulation of OTUB1 and
GRAIL expression.
GENE STRUCTURE
Soares et al. (2004) determined that the OTB1 gene contains 7 exons.
BIOCHEMICAL FEATURES
- Crystal Structure
Wiener et al. (2012) described structural and biochemical studies
elucidating how OTUB1 inhibits UBC13 and other E2 enzymes. They
unexpectedly found that OTUB1 binding to UBC13-Ub (the ubiquitin
thiolester) is allosterically regulated by free ubiquitin, which binds
to a second site in OTUB1 and increases its affinity for UBC13-Ub, while
at the same time disrupting interactions with UEV1A (602995) in a manner
that depends on the OTUB1 N terminus. Crystal structures of an
OTUB1-UBC13 complex and of OTUB1 bound to ubiquitin aldehyde and a
chemical UBC13-Ub conjugate showed that binding of free ubiquitin to
OTUB1 triggers conformational changes in the OTU domain and formation of
a ubiquitin-binding helix in the N terminus, thus promoting binding of
the conjugated donor ubiquitin in UBC13-Ub to OTUB1. The donor ubiquitin
thus cannot interact with the E2 enzyme, which has been shown to be
important for ubiquitin transfer. The N-terminal helix of OTUB1 is
positioned to interfere with UEV1A binding to UBC13, as well as with
attack on the thiolester by an acceptor ubiquitin, thereby inhibiting
K63Ub synthesis. OTUB1 binding also occludes the RING E3 binding site on
UBC13, thus providing a further component of inhibition. Wiener et al.
(2012) concluded that the general features of the inhibition mechanism
explained how OTUB1 inhibits other E2 enzymes in a noncatalytic manner.
MAPPING
By genomic sequence analysis, Balakirev et al. (2003) mapped the OTUB1
gene to chromosome 11q13.1.
*FIELD* RF
1. Balakirev, M. Y.; Tcherniuk, S. O.; Jaquinod, M.; Chroboczek, J.
: Otubains: a new family of cysteine proteases in the ubiquitin pathway. EMBO
Reports 4: 517-522, 2003.
2. Lin, J. T.; Lineberry, N. B.; Kattah, M. G.; Su, L. L.; Utz, P.
J.; Fathman, C. G.; Wu, L.: Naive CD4 T cell proliferation is controlled
by mammalian target of rapamycin regulation of GRAIL expression. J.
Immun. 182: 5919-5928, 2009.
3. Nakada, S.; Tai, I.; Panier, S.; Al-Hakim, A.; Iemura, S.; Juang,
Y.-C.; O'Donnell, L.; Kumakubo, A.; Munro, M.; Sicheri, F.; Gingras,
A.-C.; Natsume, T.; Suda, T.; Durocher, D.: Non-canonical inhibition
of DNA damage-dependent ubiquitination by OTUB1. Nature 466: 941-946,
2010.
4. Soares, L.; Seroogy, C.; Skrenta, H.; Anandasabapathy, N.; Lovelace,
P.; Chung, C. D.; Engleman, E.; Fathman, C. G.: Two isoforms of otubain
1 regulate T cell anergy via GRAIL. Nature Immun. 5: 45-54, 2004.
5. Wiener, R.; Zhang, X.; Wang, T.; Wolberger, C.: The mechanism
of OTUB1-mediated inhibition of ubiquitination. Nature 483: 618-622,
2012.
*FIELD* CN
Ada Hamosh - updated: 5/15/2012
Paul J. Converse - updated: 2/28/2011
Ada Hamosh - updated: 9/21/2010
*FIELD* CD
Paul J. Converse: 12/11/2003
*FIELD* ED
alopez: 05/16/2012
alopez: 5/16/2012
terry: 5/15/2012
mgross: 3/1/2011
terry: 2/28/2011
alopez: 9/22/2010
terry: 9/21/2010
alopez: 5/6/2008
alopez: 2/18/2004
mgross: 12/11/2003