Ubiquitin Chain Editing Revealed By Polyubiquitin Linkage-Specific Antibodies

Journal Article · Fri May 22 04:00:00 EDT 2009 · Cell 134:668,2008

OSTI ID:953026

Newton, K; Matsumoto, M L; Wertz, I E; Kirkpatrick, D S; Lill, J R; Tan, J; Dugger, D; Gordon, N; Sidhu, S S; Fellouse, F A; Komuves, L; French, D M; Ferrando, R E; Lam, C; Compaan, D; Yu, C; Bosanac, I; Hymowitz, S G; Kelley, R F; Dixit, V M

Posttranslational modification of proteins with polyubiquitin occurs in diverse signaling pathways and is tightly regulated to ensure cellular homeostasis. Studies employing ubiquitin mutants suggest that the fate of polyubiquitinated proteins is determined by which lysine within ubiquitin is linked to the C terminus of an adjacent ubiquitin. We have developed linkage-specific antibodies that recognize polyubiquitin chains joined through lysine 63 (K63) or 48 (K48). A cocrystal structure of an anti-K63 linkage Fab bound to K63-linked diubiquitin provides insight into the molecular basis for specificity. We use these antibodies to demonstrate that RIP1, which is essential for tumor necrosis factor-induced NF-{kappa}B activation, and IRAK1, which participates in signaling by interleukin-1{beta} and Toll-like receptors, both undergo polyubiquitin editing in stimulated cells. Both kinase adaptors initially acquire K63-linked polyubiquitin, while at later times K48-linked polyubiquitin targets them for proteasomal degradation. Polyubiquitin editing may therefore be a general mechanism for attenuating innate immune signaling.

• Research Organization: Stanford Linear Accelerator Center (SLAC)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC02-76SF00515
• OSTI ID: 953026
• Report Number(s): SLAC-REPRINT-2009-258
• Journal Information: Cell 134:668,2008, Journal Name: Cell 134:668,2008 Journal Issue: 4 Vol. 134; ISSN 0092-8674; ISSN CELLB5
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
ANTIBODIES
HOMEOSTASIS
LYSINE
MODIFICATIONS
MUTANTS
NECROSIS
NEOPLASMS
Other
OTHER
PHOSPHOTRANSFERASES
PROTEINS
SPECIFICITY
TARGETS