SPATA2-Mediated Binding of CYLD to HOIP Enables CYLD Recruitment to Signaling Complexes

Sebastian Kupka,Diego De Miguel,Peter Draber,Luigi Martino,Silvia Surinova,Katrin Rittinger,Henning Walczak

doi:10.1016/j.celrep.2016.07.086

Sebastian Kupka, Diego De Miguel + Show 5 more

Open Access

https://doi.org/10.1016/j.celrep.2016.07.086

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Abstract

SummaryRecruitment of the deubiquitinase CYLD to signaling complexes is mediated by its interaction with HOIP, the catalytically active component of the linear ubiquitin chain assembly complex (LUBAC). Here, we identify SPATA2 as a constitutive direct binding partner of HOIP that bridges the interaction between CYLD and HOIP. SPATA2 recruitment to TNFR1- and NOD2-signaling complexes is dependent on HOIP, and loss of SPATA2 abolishes CYLD recruitment. Deficiency in SPATA2 exerts limited effects on gene activation pathways but diminishes necroptosis induced by tumor necrosis factor (TNF), resembling loss of CYLD. In summary, we describe SPATA2 as a previously unrecognized factor in LUBAC-dependent signaling pathways that serves as an adaptor between HOIP and CYLD, thereby enabling recruitment of CYLD to signaling complexes.

Highlights

Over the past few years, the balancing roles of E3 ubiquitin ligases (E3s) and deubiquitinases (DUBs) in creating and degrading ubiquitin chains, respectively, have emerged as crucial at regulating innate and adaptive immune responses (Fiil and Gyrd-Hansen, 2014; Zinngrebe et al, 2014)
SPATA2 Is a Component of the TNF receptor 1 (TNFR1)-signaling complexes (SCs) To address whether there may be factors, in addition to CYLD, that are constitutively associated with linear ubiquitin chain assembly complex (LUBAC) and recruited to the TNFR1-SC, we performed two different mass spectrometry (MS) analyses using a modified tandem affinity purification (TAP) approach
SPATA2 was a candidate for a constitutive interaction partner of LUBAC and for a previously unrecognized component of the TNFR1-SC

Summary

Introduction

Over the past few years, the balancing roles of E3 ubiquitin ligases (E3s) and deubiquitinases (DUBs) in creating and degrading ubiquitin chains, respectively, have emerged as crucial at regulating innate and adaptive immune responses (Fiil and Gyrd-Hansen, 2014; Zinngrebe et al, 2014). Tumor necrosis factor (TNF) binding to TNF receptor 1 (TNFR1) triggers formation of the TNFR1 signaling complex (TNFR1-SC) (Walczak et al, 2012) Signals initiated from this complex result in two very different outcomes: (1) induction of gene activation via NF-kB and mitogen-activated protein (MAP) kinases and (2) induction of cell death, which can either be apoptotic or necroptotic. In the absence of LUBAC, the lack of linear ubiquitin chains in the TNFR1-SC results in defective recruitment of various components and complex destabilization (Haas et al, 2009) This shifts the signaling toward enhanced formation of a secondary SC, which induces cell death (Peltzer et al, 2014), referred to as complex II of TNFR1 signaling (Newton and Manning, 2016). Linear and other ubiquitin linkages are removed by the DUB CYLD, a process that is crucial to enable the formation of complex II, as CYLD- deficient cells are resistant to TNF-induced cell death (Draber et al, 2015; Moquin et al, 2013)

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