Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research
Date (from‐to) : 2013/04 -2015/03
Author : NAKATSUKASA Kunio
Inactivation of Cdc48p/p97 stalled retrotranslocation and triggered formation of a complex that contains the 26S proteasome, Cdc48p/p97, ubiquitinated substrates, select components of the Hrd1 complex, and the lumenal recognition factor, Yos9p. We proposed that the actions of Cdc48p/p97 and the proteasome are tightly coupled during ERAD. Our data also support a model in which the Hrd1 complex links substrate recognition and degradation on opposite sides of the ER membrane. Based on these results, we performed in vivo site-specific crosslinking experiments. Non-natural photo-reactive amino acids were incorporated into Hrd3 in yeast to detect cross-linked Hrd1. Although we could detect some cross-linked products with Hrd3, it is currently unclear if these are Hrd1-Hrd3 crosslinked products. We are now trying to scale up the system to better detect Hrd1. In addition, we are analyzing a role of the transmembrane domain of Hrd3 especially under the stress conditions.