Dominant negative mutations in yeast Hsp90 indicate triage decision mechanism targeting client proteins for degradation.

ABSTRACT: Dominant negative (DN) mutations provide valuable tools for investigating protein mechanisms but can be difficult to isolate because of their toxic effects. We used a mutational scanning approach to identify DN mutations in yeast Hsp90. In a previous mutational scan of the ATPase domain of Hsp90, we noticed that many mutations were at very low frequency after outgrowth in cells coexpressing wildtype Hsp90. Most of these depleted variants were located at the hinge of a lid that closes over ATP. To quantify toxic effects in the hinge regions, we performed mutational scanning using an inducible promoter and identified 113 variants with strong toxic effects. We analyzed individual DN mutations in detail and found that addition of the E33A mutation that prevents ATP hydrolysis by Hsp90 abrogate

SUBMITTER: Flynn JM 

PROVIDER: S-EPMC11742116 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature