About this item:

89 Views | 29 Downloads

Author Notes:

Jun Yin, junyin@gsu.edu; Yury O. Chernoff, yury.chernoff@biology.gatech.edu

B.Z., N.S., Y.O.C. and J.Y. conceived the overall idea and designed the experiments. Y.W., S.F., G.C., R.G., L.Z., D.D. and N.S. performed experiments. T.A.C., M.L., and H.K. provided technical guidance and/or intellectual input. Y.W., S.F., G.C., R.G., B.Z., N.S., Y.O.C. and J.Y. analyzed data, interpreted results, drafted and edited the manuscript. B.Z., N.S., Y.O.C. and J.Y. acquired funding. All authors reviewed and approved the final version of the manuscript.

We thank Lu Zhu and Scott D. Emr (Cornell University), David J. Katzmann (Mayo Clinic College of Medicine), Rosine Haguenauer-Tsapis (Institut Jacques Monod, France) and Pierre Morsomme (Louvain Institute of Biomolecular Science and Technology, Belgium) for kindly sharing materials and helpful discussions. We also thank Milo Fasken and Anita Corbett (Emory University) for technical advice.

The authors declare no competing interests.

Subjects:

Research Funding:

This work was supported by grants from National Natural Science Foundation of China (31770921 and 31971187 to B.Z.), NIH (R01GM114308 to N. S. and R01GM104498 to J.Y.), and NSF (1817976 to Y. O. C. and 1710460 to J.Y.).

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biochemistry & Molecular Biology
  • CLATHRIN-MEDIATED ENDOCYTOSIS
  • RNA-POLYMERASE-II
  • SACCHAROMYCES-CEREVISIAE
  • HEXOSE TRANSPORTERS
  • GENE-EXPRESSION
  • PROMOTER SYSTEM
  • PROTEIN LIGASE
  • MESSENGER-RNA
  • RIBOSOMAL-RNA
  • PSI+ PRION

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

Show all authors Show less authors

Journal Title:

CELL CHEMICAL BIOLOGY

Volume:

Volume 28, Number 9

Publisher:

, Pages 1283-+

Type of Work:

Article | Post-print: After Peer Review

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, which 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.
Export to EndNote