Distribution of Proteins at the Inner Nuclear Membrane Is Regulated by the Asi1 E3 Ligase in Saccharomyces cerevisiae

Genetics: A Periodical Record of Investigations Bearing on Heredity and Variation

Jennifer M. Gardner^1¹  Sarah E. Smith^1²  Christine J. Smoyer^1³ 

[1] Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66160^2;Stowers Institute for Medical Research, Kansas City, Missouri 64110;Stowers Institute for Medical Research, Kansas City, Missouri 64110^1

关键词: inner nuclear membrane;    ERAD;    split-GFP;    Asi1;    Pom33;    NPC;   

DOI  :  10.1534/genetics.119.301911

学科分类：医学（综合）

来源: Genetics Society of America

【 摘 要 】

Inner nuclear membrane (INM) protein composition regulates nuclear function, affecting processes such as gene expression, chromosome organization, nuclear shape, and stability. Mechanisms that drive changes in the INM proteome are poorly understood, in part because it is difficult to definitively assay INM composition rigorously and systematically. Using a split-GFP complementation system to detect INM access, we examined the distribution of all C-terminally tagged Saccharomyces cerevisiae membrane proteins in wild-type cells and in mutants affecting protein quality control pathways, such as INM-associated degradation (INMAD), ER-associated degradation, and vacuolar proteolysis. Deletion of the E3 ligase Asi1 had the most specific effect on the INM compared to mutants in vacuolar or ER-associated degradation pathways, consistent with a role for Asi1 in the INMAD pathway. Our data suggest that Asi1 not only removes mistargeted proteins at the INM, but also controls the levels and distribution of native INM components, such as the membrane nucleoporin Pom33 . Interestingly, loss of Asi1 does not affect Pom33 protein levels but instead alters Pom33 distribution in the nuclear envelope through Pom33 ubiquitination, which drives INM redistribution. Taken together, our data demonstrate that the Asi1 E3 ligase has a novel function in INM protein regulation in addition to protein turnover.

【 授权许可】

CC BY