Dataset Information

Regulation of the Endocytosis and Prion Chaperoning Machineries by Yeast E3 Ubiquitin Ligase Rsp5 as Revealed by Orthogonal Ubiquitin Transfer

ABSTRACT: Attachment of the ubiquitin (UB) peptide to proteins via the E1-E2-E3 enzymatic machinery regulates diverse biological pathways, yet identification of the substrates of E3 UB ligases remains a challenge. We overcame this challenge by constructing an “orthogonal UB transfer (OUT) cascade with yeast E3 Rsp5 to enable the exclusive delivery of an engineered UB (xUB) to Rsp5 and its substrate proteins. The OUT screen uncovered new Rsp5 substrates in yeast, such as Pal1 and Pal2 that are partners of endocytic protein Ede1, and chaperones Hsp70-Ssb, Hsp82, and Hsp104 that counteract protein misfolding and control self-perpetuating amyloid aggregates (prions), resembling those involved in human amyloid diseases. We showed that prion formation and effect of Hsp104 on prion propagation are modulated by Rsp5. Overall, our work demonstrates the capacity of OUT to deconvolute the complex E3-substrate relationships in crucial biological processes such as endocytosis and protein assembly disorders through protein ubiquitination.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Duc Duong

LAB HEAD: Jun Yin

PROVIDER: PXD023688 | Pride | 2022-10-13

REPOSITORIES: Pride

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Publications

Regulation of the endocytosis and prion-chaperoning machineries by yeast E3 ubiquitin ligase Rsp5 as revealed by orthogonal ubiquitin transfer.

Wang Yiyang Y Wang Yiyang Y Fang Shuai S Chen Geng G Ganti Rakhee R Chernova Tatiana A TA Zhou Li L Duong Duc D Kiyokawa Hiroaki H Li Ming M Zhao Bo B Shcherbik Natalia N Chernoff Yury O YO Yin Jun J

Cell chemical biology 20210304 9

Attachment of the ubiquitin (UB) peptide to proteins via the E1-E2-E3 enzymatic machinery regulates diverse biological pathways, yet identification of the substrates of E3 UB ligases remains a challenge. We overcame this challenge by constructing an "orthogonal UB transfer" (OUT) cascade with yeast E3 Rsp5 to enable the exclusive delivery of an engineered UB (xUB) to Rsp5 and its substrate proteins. The OUT screen uncovered new Rsp5 substrates in yeast, such as Pal1 and Pal2, which are partners ...[more]

PMID: 33667410

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Dataset Information

Regulation of the Endocytosis and Prion Chaperoning Machineries by Yeast E3 Ubiquitin Ligase Rsp5 as Revealed by Orthogonal Ubiquitin Transfer

Publications

Regulation of the endocytosis and prion-chaperoning machineries by yeast E3 ubiquitin ligase Rsp5 as revealed by orthogonal ubiquitin transfer.

Similar Datasets

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