The p62 scaffold regulates nerve growth factor-induced NF-kappaB activation by influencing TRAF6 polyubiquitination.
Sequestosome 1/p62 is a scaffolding protein with several interaction modules that include a PB1 dimerization domain, a TRAF6 (tumor necrosis factor receptor-associated factor 6) binding site, and a ubiquitin-associating (UBA) domain. Here, we report that p62 functions to facilitate K63-polyubiquitination of TRAF6 and thereby mediates nerve growth factor-induced activation of the NF-kappaB pathway. In brain of p62 knock-out mice we did not recover polyubiquitinated TRAF6. The UBA domain binds polyubiquitin chains and deletion of p62-UBA domain or mutation of F406V within the ubiquitin binding pocket of the UBA domain abolished TRAF6 polyubiquitination. Likewise, deletion of p62 N-terminal dimerization domain or the TRAF6 binding site had similar effects on both polyubiquitination and oligomerization of TRAF6. Nerve growth factor treatment of PC12 cells induced TRAF6 polyubiquitination along with formation of a p62-TRAF6-IKKbeta-PKC iota signal complex, while inhibition of the p62/TRAF6 interaction had an opposite effect. These results provide evidence for a mechanism whereby p62 serves to regulate the NF-kappaB pathway.
Wooten
Marie W
MW
Department of Biological Sciences, Program in Cell and Molecular Biosciences, Auburn University, Alabama 36849, USA. wootemw@auburn.edu
Geetha
Thangiah
T
Seibenhener
M Lamar
ML
Babu
J Ramesh
JR
Diaz-Meco
Maria T
MT
Moscat
Jorge
J
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D017353:Gene Deletion; D005990:Glycerol / Q000737:chemistry; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D047468:Immunoprecipitation; D051379:Mice; D018345:Mice, Knockout; D008957:Models, Genetic; D016328:NF-kappa B / Q000378:metabolism*; D020932:Nerve Growth Factor / Q000378:metabolism; D016716:PC12 Cells; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000502:physiology*; D051381:Rats; D000071456:Sequestosome-1 Protein; D015398:Signal Transduction; D048029:TNF Receptor-Associated Factor 6 / Q000378:metabolism*; D013997:Time Factors; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism*; D014461:Ultracentrifugation
D048868:Adaptor Proteins, Signal Transducing; D006360:Heat-Shock Proteins; D016328:NF-kappa B; D011506:Proteins; C099718:SQSTM1 protein, human; D000071456:Sequestosome-1 Protein; C491079:Sqstm1 protein, rat; D048029:TNF Receptor-Associated Factor 6; D025801:Ubiquitin; D020932:Nerve Growth Factor; D005990:Glycerol