ID   OTUL_MOUSE              Reviewed;         352 AA.
AC   Q3UCV8; Q3UY59; Q5M8N1; Q8R027;
DT   28-NOV-2006, integrated into UniProtKB/Swiss-Prot.
DT   11-OCT-2005, sequence version 1.
DT   24-JAN-2024, entry version 116.
DE   RecName: Full=Ubiquitin thioesterase otulin {ECO:0000305};
DE            EC=3.4.19.12 {ECO:0000269|PubMed:23708998, ECO:0000269|PubMed:29950720};
DE   AltName: Full=Deubiquitinating enzyme otulin {ECO:0000250|UniProtKB:Q96BN8};
DE   AltName: Full=OTU domain-containing deubiquitinase with linear linkage specificity {ECO:0000250|UniProtKB:Q96BN8};
DE   AltName: Full=Ubiquitin thioesterase Gumby {ECO:0000303|PubMed:23708998};
GN   Name=Otulin {ECO:0000312|MGI:MGI:3577015};
GN   Synonyms=Fam105b {ECO:0000312|MGI:MGI:3577015}, Gum
GN   {ECO:0000303|PubMed:23708998};
OS   Mus musculus (Mouse).
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC   Murinae; Mus; Mus.
OX   NCBI_TaxID=10090;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA].
RC   STRAIN=C57BL/6J; TISSUE=Bone marrow, and Olfactory bulb;
RX   PubMed=16141072; DOI=10.1126/science.1112014;
RA   Carninci P., Kasukawa T., Katayama S., Gough J., Frith M.C., Maeda N.,
RA   Oyama R., Ravasi T., Lenhard B., Wells C., Kodzius R., Shimokawa K.,
RA   Bajic V.B., Brenner S.E., Batalov S., Forrest A.R., Zavolan M., Davis M.J.,
RA   Wilming L.G., Aidinis V., Allen J.E., Ambesi-Impiombato A., Apweiler R.,
RA   Aturaliya R.N., Bailey T.L., Bansal M., Baxter L., Beisel K.W., Bersano T.,
RA   Bono H., Chalk A.M., Chiu K.P., Choudhary V., Christoffels A.,
RA   Clutterbuck D.R., Crowe M.L., Dalla E., Dalrymple B.P., de Bono B.,
RA   Della Gatta G., di Bernardo D., Down T., Engstrom P., Fagiolini M.,
RA   Faulkner G., Fletcher C.F., Fukushima T., Furuno M., Futaki S.,
RA   Gariboldi M., Georgii-Hemming P., Gingeras T.R., Gojobori T., Green R.E.,
RA   Gustincich S., Harbers M., Hayashi Y., Hensch T.K., Hirokawa N., Hill D.,
RA   Huminiecki L., Iacono M., Ikeo K., Iwama A., Ishikawa T., Jakt M.,
RA   Kanapin A., Katoh M., Kawasawa Y., Kelso J., Kitamura H., Kitano H.,
RA   Kollias G., Krishnan S.P., Kruger A., Kummerfeld S.K., Kurochkin I.V.,
RA   Lareau L.F., Lazarevic D., Lipovich L., Liu J., Liuni S., McWilliam S.,
RA   Madan Babu M., Madera M., Marchionni L., Matsuda H., Matsuzawa S., Miki H.,
RA   Mignone F., Miyake S., Morris K., Mottagui-Tabar S., Mulder N., Nakano N.,
RA   Nakauchi H., Ng P., Nilsson R., Nishiguchi S., Nishikawa S., Nori F.,
RA   Ohara O., Okazaki Y., Orlando V., Pang K.C., Pavan W.J., Pavesi G.,
RA   Pesole G., Petrovsky N., Piazza S., Reed J., Reid J.F., Ring B.Z.,
RA   Ringwald M., Rost B., Ruan Y., Salzberg S.L., Sandelin A., Schneider C.,
RA   Schoenbach C., Sekiguchi K., Semple C.A., Seno S., Sessa L., Sheng Y.,
RA   Shibata Y., Shimada H., Shimada K., Silva D., Sinclair B., Sperling S.,
RA   Stupka E., Sugiura K., Sultana R., Takenaka Y., Taki K., Tammoja K.,
RA   Tan S.L., Tang S., Taylor M.S., Tegner J., Teichmann S.A., Ueda H.R.,
RA   van Nimwegen E., Verardo R., Wei C.L., Yagi K., Yamanishi H.,
RA   Zabarovsky E., Zhu S., Zimmer A., Hide W., Bult C., Grimmond S.M.,
RA   Teasdale R.D., Liu E.T., Brusic V., Quackenbush J., Wahlestedt C.,
RA   Mattick J.S., Hume D.A., Kai C., Sasaki D., Tomaru Y., Fukuda S.,
RA   Kanamori-Katayama M., Suzuki M., Aoki J., Arakawa T., Iida J., Imamura K.,
RA   Itoh M., Kato T., Kawaji H., Kawagashira N., Kawashima T., Kojima M.,
RA   Kondo S., Konno H., Nakano K., Ninomiya N., Nishio T., Okada M., Plessy C.,
RA   Shibata K., Shiraki T., Suzuki S., Tagami M., Waki K., Watahiki A.,
RA   Okamura-Oho Y., Suzuki H., Kawai J., Hayashizaki Y.;
RT   "The transcriptional landscape of the mammalian genome.";
RL   Science 309:1559-1563(2005).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=C57BL/6J;
RX   PubMed=19468303; DOI=10.1371/journal.pbio.1000112;
RA   Church D.M., Goodstadt L., Hillier L.W., Zody M.C., Goldstein S., She X.,
RA   Bult C.J., Agarwala R., Cherry J.L., DiCuccio M., Hlavina W., Kapustin Y.,
RA   Meric P., Maglott D., Birtle Z., Marques A.C., Graves T., Zhou S.,
RA   Teague B., Potamousis K., Churas C., Place M., Herschleb J., Runnheim R.,
RA   Forrest D., Amos-Landgraf J., Schwartz D.C., Cheng Z., Lindblad-Toh K.,
RA   Eichler E.E., Ponting C.P.;
RT   "Lineage-specific biology revealed by a finished genome assembly of the
RT   mouse.";
RL   PLoS Biol. 7:E1000112-E1000112(2009).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA] OF 106-352.
RC   TISSUE=Mammary gland, and Molar;
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 project:
RT   the Mammalian Gene Collection (MGC).";
RL   Genome Res. 14:2121-2127(2004).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, DISRUPTION PHENOTYPE,
RP   DEVELOPMENTAL STAGE, ACTIVE SITE, INTERACTION WITH RNF31 AND DVL2, AND
RP   MUTAGENESIS OF TRP-96; CYS-129; ASP-336 AND 349-GLU--VAL-352.
RX   PubMed=23708998; DOI=10.1038/nature12296;
RA   Rivkin E., Almeida S.M., Ceccarelli D.F., Juang Y.C., Maclean T.A.,
RA   Srikumar T., Huang H., Dunham W.H., Fukumura R., Xie G., Gondo Y.,
RA   Raught B., Gingras A.C., Sicheri F., Cordes S.P.;
RT   "The linear ubiquitin-specific deubiquitinase gumby regulates
RT   angiogenesis.";
RL   Nature 498:318-324(2013).
RN   [5]
RP   FUNCTION, AND DISRUPTION PHENOTYPE.
RX   PubMed=27523608; DOI=10.1016/j.cell.2016.07.019;
RA   Damgaard R.B., Walker J.A., Marco-Casanova P., Morgan N.V.,
RA   Titheradge H.L., Elliott P.R., McHale D., Maher E.R., McKenzie A.N.,
RA   Komander D.;
RT   "The deubiquitinase OTULIN is an essential negative regulator of
RT   inflammation and autoimmunity.";
RL   Cell 166:1215-1230(2016).
RN   [6]
RP   FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF CYS-129.
RX   PubMed=29950720; DOI=10.1038/s41586-018-0256-2;
RA   Heger K., Wickliffe K.E., Ndoja A., Zhang J., Murthy A., Dugger D.L.,
RA   Maltzman A., de Sousa E Melo F., Hung J., Zeng Y., Verschueren E.,
RA   Kirkpatrick D.S., Vucic D., Lee W.P., Roose-Girma M., Newman R.J.,
RA   Warming S., Hsiao Y.C., Komuves L.G., Webster J.D., Newton K., Dixit V.M.;
RT   "OTULIN limits cell death and inflammation by deubiquitinating LUBAC.";
RL   Nature 559:120-124(2018).
CC   -!- FUNCTION: Deubiquitinase that specifically removes linear ('Met-1'-
CC       linked) polyubiquitin chains to substrates and acts as a regulator of
CC       angiogenesis and innate immune response (PubMed:23708998,
CC       PubMed:27523608, PubMed:29950720). Required during angiogenesis,
CC       craniofacial and neuronal development by regulating the canonical Wnt
CC       signaling together with the LUBAC complex (PubMed:23708998). Acts as a
CC       negative regulator of NF-kappa-B by regulating the activity of the
CC       LUBAC complex (By similarity). OTULIN function is mainly restricted to
CC       homeostasis of the LUBAC complex: acts by removing 'Met-1'-linked
CC       autoubiquitination of the LUBAC complex, thereby preventing
CC       inactivation of the LUBAC complex (PubMed:29950720). Acts as a key
CC       negative regulator of inflammation by restricting spontaneous
CC       inflammation and maintaining immune homeostasis (PubMed:27523608,
CC       PubMed:29950720). In myeloid cell, required to prevent unwarranted
CC       secretion of cytokines leading to inflammation and autoimmunity by
CC       restricting linear polyubiquitin formation (PubMed:27523608). Plays a
CC       role in innate immune response by restricting linear polyubiquitin
CC       formation on LUBAC complex in response to NOD2 stimulation, probably to
CC       limit NOD2-dependent pro-inflammatory signaling (By similarity).
CC       {ECO:0000250|UniProtKB:Q96BN8, ECO:0000269|PubMed:23708998,
CC       ECO:0000269|PubMed:27523608, ECO:0000269|PubMed:29950720}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Thiol-dependent hydrolysis of ester, thioester, amide, peptide
CC         and isopeptide bonds formed by the C-terminal Gly of ubiquitin (a 76-
CC         residue protein attached to proteins as an intracellular targeting
CC         signal).; EC=3.4.19.12; Evidence={ECO:0000269|PubMed:23708998,
CC         ECO:0000269|PubMed:29950720};
CC   -!- SUBUNIT: Interacts (via the PUB domain) with RNF31 (via the PIM motif);
CC       the interaction is direct (PubMed:23708998). Interacts with DVL2
CC       (PubMed:23708998). {ECO:0000269|PubMed:23708998}.
CC   -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:23708998}.
CC   -!- DEVELOPMENTAL STAGE: Enriched in a subset of endothelial cells near
CC       presumptive tips of vessels and vascular buds (at protein level).
CC       {ECO:0000269|PubMed:23708998}.
CC   -!- DOMAIN: The specificity for linear polyubiquitin is given by the 'Glu-
CC       16' residue in ubiquitin chain. {ECO:0000250|UniProtKB:Q96BN8}.
CC   -!- DOMAIN: The PIM (PUB-interaction motif) motif mediates interaction with
CC       the PUB domain of RNF31. Does not interact with other PUB domain-
CC       containing proteins. Phosphorylation at Tyr-56 prevents interaction
CC       with RNF31. {ECO:0000250|UniProtKB:Q96BN8}.
CC   -!- PTM: Ubiquitinated. {ECO:0000250|UniProtKB:Q96BN8}.
CC   -!- PTM: Acetylated. {ECO:0000250|UniProtKB:Q96BN8}.
CC   -!- PTM: Phosphorylated. Phosphorylation at Tyr-56 prevents interaction
CC       with RNF31; dephosphorylation promotes interaction with RNF31 and the
CC       LUBAC complex. {ECO:0000250|UniProtKB:Q96BN8}.
CC   -!- DISRUPTION PHENOTYPE: Embryonic lethality (PubMed:23708998,
CC       PubMed:27523608). Specific deletion in immune cells leads to acute
CC       systemic inflammation characterized by rapid weight loss, increased
CC       levels of pro-inflammatory cytokines in serum, neutrophilia with all
CC       the hallmarks of emergency granulopoiesis (PubMed:27523608). Specific
CC       deletion in T- or B-cells generates healthy mice with no overt
CC       inflammatory phenotypes (PubMed:27523608). In contrast, specific
CC       deletion in myeloid cells results in a strong inflammatory phenotype,
CC       characterized by chronic inflammation and autoimmunity, caused by
CC       sterile autoactivation of inflammatory pathways (PubMed:27523608).
CC       {ECO:0000269|PubMed:23708998, ECO:0000269|PubMed:27523608}.
CC   -!- SIMILARITY: Belongs to the peptidase C65 family. Otulin subfamily.
CC       {ECO:0000305}.
CC   -!- SEQUENCE CAUTION:
CC       Sequence=AAH87945.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
CC       Sequence=BAE22354.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
CC   ---------------------------------------------------------------------------
CC   Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC   Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC   ---------------------------------------------------------------------------
DR   EMBL; AK134955; BAE22354.1; ALT_INIT; mRNA.
DR   EMBL; AK150371; BAE29504.1; -; mRNA.
DR   EMBL; GL456173; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR   EMBL; BC028541; AAH28541.1; -; mRNA.
DR   EMBL; BC087945; AAH87945.1; ALT_INIT; mRNA.
DR   CCDS; CCDS49586.1; -.
DR   RefSeq; NP_001013814.2; NM_001013792.2.
DR   AlphaFoldDB; Q3UCV8; -.
DR   SMR; Q3UCV8; -.
DR   BioGRID; 240910; 3.
DR   STRING; 10090.ENSMUSP00000057893; -.
DR   iPTMnet; Q3UCV8; -.
DR   PhosphoSitePlus; Q3UCV8; -.
DR   SwissPalm; Q3UCV8; -.
DR   EPD; Q3UCV8; -.
DR   MaxQB; Q3UCV8; -.
DR   PaxDb; 10090-ENSMUSP00000057893; -.
DR   PeptideAtlas; Q3UCV8; -.
DR   ProteomicsDB; 294136; -.
DR   Pumba; Q3UCV8; -.
DR   Antibodypedia; 56178; 119 antibodies from 22 providers.
DR   DNASU; 432940; -.
DR   Ensembl; ENSMUST00000059662.8; ENSMUSP00000057893.8; ENSMUSG00000046034.9.
DR   GeneID; 432940; -.
DR   KEGG; mmu:432940; -.
DR   UCSC; uc007vjr.1; mouse.
DR   AGR; MGI:3577015; -.
DR   CTD; 90268; -.
DR   MGI; MGI:3577015; Otulin.
DR   VEuPathDB; HostDB:ENSMUSG00000046034; -.
DR   eggNOG; ENOG502QTB8; Eukaryota.
DR   GeneTree; ENSGT00390000009802; -.
DR   HOGENOM; CLU_051856_1_1_1; -.
DR   InParanoid; Q3UCV8; -.
DR   OMA; CRKEWRG; -.
DR   OrthoDB; 3151774at2759; -.
DR   PhylomeDB; Q3UCV8; -.
DR   TreeFam; TF328709; -.
DR   Reactome; R-MMU-5357905; Regulation of TNFR1 signaling.
DR   Reactome; R-MMU-8866652; Synthesis of active ubiquitin: roles of E1 and E2 enzymes.
DR   BioGRID-ORCS; 432940; 9 hits in 81 CRISPR screens.
DR   ChiTaRS; Otulin; mouse.
DR   PRO; PR:Q3UCV8; -.
DR   Proteomes; UP000000589; Chromosome 15.
DR   RNAct; Q3UCV8; Protein.
DR   Bgee; ENSMUSG00000046034; Expressed in yolk sac and 221 other cell types or tissues.
DR   ExpressionAtlas; Q3UCV8; baseline and differential.
DR   GO; GO:0005737; C:cytoplasm; IDA:UniProtKB.
DR   GO; GO:0004843; F:cysteine-type deubiquitinase activity; IDA:UniProtKB.
DR   GO; GO:0008234; F:cysteine-type peptidase activity; IMP:UniProtKB.
DR   GO; GO:0045087; P:innate immune response; ISS:UniProtKB.
DR   GO; GO:0050728; P:negative regulation of inflammatory response; IMP:UniProtKB.
DR   GO; GO:0032088; P:negative regulation of NF-kappaB transcription factor activity; IMP:UniProtKB.
DR   GO; GO:0070431; P:nucleotide-binding oligomerization domain containing 2 signaling pathway; ISS:UniProtKB.
DR   GO; GO:1990108; P:protein linear deubiquitination; IDA:UniProtKB.
DR   GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR   GO; GO:0060828; P:regulation of canonical Wnt signaling pathway; IDA:UniProtKB.
DR   GO; GO:0010803; P:regulation of tumor necrosis factor-mediated signaling pathway; IMP:UniProtKB.
DR   GO; GO:0002040; P:sprouting angiogenesis; IMP:UniProtKB.
DR   GO; GO:0016055; P:Wnt signaling pathway; IEA:UniProtKB-KW.
DR   CDD; cd22799; OTU_OTUL; 1.
DR   Gene3D; 3.30.200.60; Peptidase C65 Otubain, subdomain 1; 1.
DR   Gene3D; 1.20.1300.20; Peptidase C65 Otubain, subdomain 2; 1.
DR   InterPro; IPR023235; FAM105.
DR   InterPro; IPR023237; Otulin.
DR   InterPro; IPR042468; Peptidase_C65_otubain_sub1.
DR   InterPro; IPR042467; Peptidase_C65_otubain_sub2.
DR   PANTHER; PTHR33662; OTU DEUBIQUITINASE WITH LINEAR LINKAGE-SPECIFICITY A-RELATED; 1.
DR   PANTHER; PTHR33662:SF2; UBIQUITIN THIOESTERASE OTULIN; 1.
DR   Pfam; PF16218; Peptidase_C101; 1.
DR   PRINTS; PR02055; PROTEINF105.
DR   PRINTS; PR02057; PROTEINF105B.
DR   Genevisible; Q3UCV8; MM.
PE   1: Evidence at protein level;
KW   Acetylation; Angiogenesis; Coiled coil; Cytoplasm; Hydrolase; Immunity;
KW   Innate immunity; Phosphoprotein; Protease; Reference proteome;
KW   Thiol protease; Ubl conjugation; Ubl conjugation pathway;
KW   Wnt signaling pathway.
FT   CHAIN           1..352
FT                   /note="Ubiquitin thioesterase otulin"
FT                   /id="PRO_0000261638"
FT   DOMAIN          118..346
FT                   /note="OTU"
FT   REGION          1..49
FT                   /note="Disordered"
FT                   /evidence="ECO:0000256|SAM:MobiDB-lite"
FT   REGION          95..96
FT                   /note="Linear diubiquitin binding"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   REGION          124..126
FT                   /note="Linear diubiquitin binding"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   REGION          255..259
FT                   /note="Linear diubiquitin binding"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   REGION          283..289
FT                   /note="Linear diubiquitin binding"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   REGION          336..338
FT                   /note="Linear diubiquitin binding"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   COILED          49..73
FT                   /evidence="ECO:0000255"
FT   MOTIF           52..57
FT                   /note="PIM motif"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   MOTIF           349..352
FT                   /note="PDZ-binding"
FT   ACT_SITE        126
FT                   /evidence="ECO:0000269|PubMed:23708998"
FT   ACT_SITE        129
FT                   /note="Nucleophile"
FT                   /evidence="ECO:0000269|PubMed:23708998"
FT   ACT_SITE        339
FT                   /evidence="ECO:0000269|PubMed:23708998"
FT   SITE            314
FT                   /note="Linear diubiquitin binding"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   MOD_RES         56
FT                   /note="Phosphotyrosine"
FT                   /evidence="ECO:0000250|UniProtKB:Q96BN8"
FT   MUTAGEN         96
FT                   /note="W->R: In Gum(W96R) mutant; defects in embryonic
FT                   angiogenesis. Embryos appear normal before 11.5 dpc but die
FT                   between 12.5 dpc-14 dpc, probably due to defects in
FT                   organization of branching vascular networks in the head and
FT                   trunk."
FT                   /evidence="ECO:0000269|PubMed:23708998"
FT   MUTAGEN         129
FT                   /note="C->A,S: Abolishes deubiquitinase activity without
FT                   affecting interaction with RNF31. Lethality at midgestation
FT                   in knockin mice caused by inactivation of the LUBAC
FT                   complex, leading to cell death mediated by TNFR1 and
FT                   RIPK1."
FT                   /evidence="ECO:0000269|PubMed:23708998,
FT                   ECO:0000269|PubMed:29950720"
FT   MUTAGEN         336
FT                   /note="D->E: In Gum(D366E) mutant; defects in embryonic
FT                   angiogenesis. Weaker mutant compared to Gum(W96R) mutant."
FT                   /evidence="ECO:0000269|PubMed:23708998"
FT   MUTAGEN         349..352
FT                   /note="ETSV->AAAA: Does not affect interaction with RNF31."
FT                   /evidence="ECO:0000269|PubMed:23708998"
SQ   SEQUENCE   352 AA;  40320 MW;  435B9B3F4BDEEDDA CRC64;
     MSRGTMPQPG AWPGASCAET PAREAGAAAR DGGKVTAGAQ PRAATRCPAE HEEDMYRAAD
     EIEKEKELLI HERGISEPRL SVAPEMDIMD YCKKEWRGNT QKATCMKKGY EEVSQKFTSI
     RRVRGDNYCA LRATLFQAMS QLAELPPWLQ DLELILLPEK LINKYTWIKQ WKLGLKFDGK
     SEDLVEKIKE SLALLRKKWV SLAAMKTAEA RQTACDELFT NEEEEYSLYE AVKFLMLNRA
     IELYDDKEKG KEVPFFSVLL FARDTSNDPE QLLRNHLNQV GHTGGLEQVE MFLLAYAVRH
     SIRVYRLSKY NTEEFITVYP TDPPKDWPMV TLIAEDDRHY NIPVRVCEET SV
//