OTULIN antagonizes LUBAC signaling by specifically hydrolyzing met1-linked polyubiquitin

TY - JOUR

T1 - OTULIN antagonizes LUBAC signaling by specifically hydrolyzing met1-linked polyubiquitin

AU - Keusekotten, K.

AU - Elliott, P.R.

AU - Kulathu, Y.

AU - Wauer, T.

AU - Hospenthal, M.K.

AU - Komander, D.

AU - Glockner, L.

AU - Krappmann, D.

AU - Fiil, B.K.

AU - Damgaard, R.B.

AU - Gyrd-Hansen, M.

AU - Hofmann, K.

PY - 2013/6/6

Y1 - 2013/6/6

N2 - The linear ubiquitin (Ub) chain assembly complex (LUBAC) is an E3 ligase that specifically assembles Met1-linked (also known as linear) Ub chains that regulate nuclear factor κB (NF-κB) signaling. Deubiquitinases (DUBs) are key regulators of Ub signaling, but a dedicated DUB for Met1 linkages has not been identified. Here, we reveal a previously unannotated human DUB, OTULIN (also known as FAM105B), which is exquisitely specific for Met1 linkages. Crystal structures of the OTULIN catalytic domain in complex with diubiquitin reveal Met1-specific Ub-binding sites and a mechanism of substrate-assisted catalysis in which the proximal Ub activates the catalytic triad of the protease. Mutation of Ub Glu16 inhibits OTULIN activity by reducing k cat 240-fold. OTULIN overexpression or knockdown affects NF-κB responses to LUBAC, TNFα, and poly(I:C) and sensitizes cells to TNFα-induced cell death. We show that OTULIN binds LUBAC and that overexpression of OTULIN prevents TNFα-induced NEMO association with ubiquitinated RIPK1. Our data suggest that OTULIN regulates Met1-polyUb signaling.

AB - The linear ubiquitin (Ub) chain assembly complex (LUBAC) is an E3 ligase that specifically assembles Met1-linked (also known as linear) Ub chains that regulate nuclear factor κB (NF-κB) signaling. Deubiquitinases (DUBs) are key regulators of Ub signaling, but a dedicated DUB for Met1 linkages has not been identified. Here, we reveal a previously unannotated human DUB, OTULIN (also known as FAM105B), which is exquisitely specific for Met1 linkages. Crystal structures of the OTULIN catalytic domain in complex with diubiquitin reveal Met1-specific Ub-binding sites and a mechanism of substrate-assisted catalysis in which the proximal Ub activates the catalytic triad of the protease. Mutation of Ub Glu16 inhibits OTULIN activity by reducing k cat 240-fold. OTULIN overexpression or knockdown affects NF-κB responses to LUBAC, TNFα, and poly(I:C) and sensitizes cells to TNFα-induced cell death. We show that OTULIN binds LUBAC and that overexpression of OTULIN prevents TNFα-induced NEMO association with ubiquitinated RIPK1. Our data suggest that OTULIN regulates Met1-polyUb signaling.

UR - http://www.scopus.com/inward/record.url?scp=84878862687&partnerID=8YFLogxK

U2 - 10.1016/j.cell.2013.05.014

DO - 10.1016/j.cell.2013.05.014

M3 - Journal article

C2 - 23746843

AN - SCOPUS:84878862687

SN - 0092-8674

VL - 153

SP - 1312

EP - 1326

JO - Cell

JF - Cell

IS - 6

ER -