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bioRxiv · 10.1101/442970

Asi1 regulates the distribution of proteins at the inner nuclear membrane in Saccharomyces cerevisiae

Smoyer, C. J.·Smith, S. E.·McCroskey, S.·Unruh, J.·Jaspersen, S. L.
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Abstract

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 (ERAD) and vacuolar proteolysis. Deletion of the E3 ligase Asi1 had the most pronounced 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 suggests that Asi1 not only removes mis-targeted proteins at the INM, but it 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 NE 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.

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Smoyer, C. J., Smith, S. E., McCroskey, S., Unruh, J., Jaspersen, S. L.. 2018-10-14. Asi1 regulates the distribution of proteins at the inner nuclear membrane in Saccharomyces cerevisiae. https://doi.org/10.1101/442970

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