Premium
Accumulation of polyubiquitinated proteins by overexpression of RBCC protein interacting with protein kinase C2, a splice variant of ubiquitin ligase RBCC protein interacting with protein kinase C1
Author(s) -
Yoshimoto Nobuo,
Tatematsu Kenji,
Okajima Toshihide,
Tanizawa Katsuyuki,
Kuroda Shun’ichi
Publication year - 2009
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2009.07350.x
Subject(s) - immunoprecipitation , ubiquitin , biology , ubiquitin ligase , protein subunit , uniprot , protein kinase a , microbiology and biotechnology , proteasome , cop9 signalosome , biochemistry , kinase , protease , enzyme , gene , peptide hydrolases
The nuclear–cytoplasmic shuttling protein RBCC protein interacting with protein kinase C1 (RBCK1) possesses transcriptional and ubiquitin ligase activities. We have recently reported that RBCC protein interacting with protein kinase C2 (RBCK2), a RING–in‐between‐RING fingers domain‐lacking splice variant of RBCK1, lacks transcriptional activity, but rather represses the RBCK1‐mediated transcriptional activity as a cytoplasmic tethering protein for RBCK1. In this study, we have found that RBCK2 overexpressed in human embryonic kidney 293 cells interacts with the polyubiquitin chain and the polyubiquitin‐interacting subunit S5a, and significantly increases the intracellular amount of polyubiquitinated proteins. These results strongly suggested that RBCK2 functions as an adaptor protein for the polyubiquitinated protein and the S5a subunit in 26S proteasome through its novel zinc finger motif and ubiquitin‐like domain, respectively, presumably delivering the polyubiquitinated proteins to the entrance of the 26S proteasome catalytic domain for degradation. Structured digital abstract• MINT‐7261426 : S5a (uniprotkb: P55036 ) binds ( MI:0407 ) to Rbck1 (uniprotkb: Q62921 ) by pull down ( MI:0096 ) • MINT‐7261435 : S5a (uniprotkb: P55036 ) physically interacts ( MI:0915 ) with Rbck1 (uniprotkb: Q62921 ) by anti bait coimmunoprecipitation ( MI:0006 ) • MINT‐7261448 : S5a (uniprotkb: P55036 ) physically interacts ( MI:0915 ) with Rbck1 (uniprotkb: Q62921 ) by anti tag coimmunoprecipitation ( MI:0007 ) • MINT‐7261462 : Rbck1 (uniprotkb: Q62921 ) physically interacts ( MI:0915 ) with Ubiquitin (uniprotkb: P62988 ) by anti tag coimmunoprecipitation ( MI:0007 ) • MINT‐7261503 : S5a (uniprotkb: P55036 ) binds ( MI:0407 ) to Ubiquitin (uniprotkb: P62988 ) by pull down ( MI:0096 ) • MINT‐7261477 : Rbck1 (uniprotkb: Q62921 ) binds ( MI:0407 ) to Ubiquitin (uniprotkb: P62988 ) by pull down ( MI:0096 )