Lys63/Met1-hybrid ubiquitin chains are commonly formed during the activation of innate immune signalling

https://doi.org/10.1016/j.bbrc.2016.04.141Get rights and content
Under a Creative Commons license
open access

Highlights

  • Ubiquitin chains containing both Lys63 and Met1 linkages are commonly formed during innate immune signaling.

  • Lys63/Met1-hybrid chains become attached to RIP1 and RIP2 in the TNFR1 and NOD1 signaling networks, respectively.

  • Potential advantages of Lys63/Met1-hybrids over separate ubiquitin chains are proposed.

  • Met1-linked ubiquitin is attached to TNFR1 without formation of a hybrid ubiquitin chain.

Abstract

We have reported previously that activation of the MyD88-signaling network rapidly induces the formation of hybrid ubiquitin chains containing both Lys63-linked and Met1-linked ubiquitin (Ub) oligomers, some of which are attached covalently to Interleukin Receptor Associated kinase 1. Here we show that Lys63/Met1-Ub hybrids are also formed rapidly when the TNFR1/TRADD, TLR3/TRIF- and NOD1/RIP2-signaling networks are activated, some of which are attached covalently to Receptor-Interacting Protein 1 (TNFR1 pathway) or Receptor-Interacting Protein 2 (NOD1 pathway). These observations suggest that the formation of Lys63/Met1-Ub hybrids are of general significance for the regulation of innate immune signaling systems, and their potential roles in vivo are discussed. We also report that TNFα induces the attachment of Met1-linked Ub chains directly to TNF receptor 1, which do not seem to be attached covalently to Lys63-linked or other types of ubiquitin chain.

Keywords

Ubiquitin
Innate immunity
TNF
TLR3
NOD1
LUBAC

Abbreviations

AMSH-LP
AMSH-like protein
λ-PPase
bacteriophage λ protein phosphatase
BMDM
bone marrow-derived macrophages
JNK
c-Jun N-terminal kinase
cIAP
cellular Inhibitor of Apoptosis
DUB
deubiquitylase
dsRNA
double-stranded RNA
GST
glutathione-S-transferase
HRP
horseradish peroxidase
IKK
IκB kinase
IL-1
interleukin-1
IL-1R
IL-1 receptor
IRAK
IL-1R-Associated Kinase
LUBAC
Linear UBiquitin Assembly Complex
K63-Ub
Lys63-linked ubiquitin
M-CSF
Macrophage Colony Stimulating Factor
M1-Ub
Met1-linked ubiquitin
MyD88
Myeloid Differentiation primary response gene 88
NEMO
NF-κB Essential Modifier
NOD
Nucleotide Oligomerisation Domain
PRRs
Pathogen Recognition Receptors
PNGase F
Protein Asparaginyl Glycosidase F
RIP
Receptor-Interacting Protein
TAK1
TGFβ-activated kinase 1
TAB
TAK1-binding protein
TRAF
TNF-Receptor-Associated Factor
TLR
Toll-Like Receptor
TRIF
TIR-domain-containing adapter-inducing interferon-β
TNFα
Tumour Necrosis Factor α
TNFR1
TNFα Receptor 1
TRADD
TNF-receptor associated death domain protein
Ub
Ubiquitin
USP
Ub-specific protease

Cited by (0)

1

Joint First Authors.