Acetylation of the yeast Hsp40 chaperone protein Ydj1 fine-tunes proteostasis and translational fidelity

PLoS Genet. 2024 Dec 9;20(12):e1011338. doi: 10.1371/journal.pgen.1011338. eCollection 2024 Dec.

Abstract

Proteostasis, the maintenance of cellular protein balance, is essential for cell viability and is highly conserved across all organisms. Newly synthesized proteins, or "clients," undergo sequential processing by Hsp40, Hsp70, and Hsp90 chaperones to achieve proper folding and functionality. Despite extensive characterization of post-translational modifications (PTMs) on Hsp70 and Hsp90, the modifications on Hsp40 remain less understood. This study aims to elucidate the role of lysine acetylation on the yeast Hsp40, Ydj1. By mutating acetylation sites on Ydj1's J-domain to either abolish or mimic constitutive acetylation, we observed that preventing acetylation had no noticeable phenotypic impact, whereas acetyl-mimic mutants exhibited various defects indicative of impaired Ydj1 function. Proteomic analysis revealed several Ydj1 interactions affected by J-domain acetylation, notably with proteins involved in translation. Further investigation uncovered a novel role for Ydj1 acetylation in stabilizing ribosomal subunits and ensuring translational fidelity. Our data suggest that acetylation may facilitate the transfer of Ydj1 between Ssa1 and Hsp82. Collectively, this work highlights the critical role of Ydj1 acetylation in proteostasis and translational fidelity.

MeSH terms

  • Acetylation
  • Adenosine Triphosphatases / genetics
  • Adenosine Triphosphatases / metabolism
  • HSP40 Heat-Shock Proteins* / genetics
  • HSP40 Heat-Shock Proteins* / metabolism
  • HSP70 Heat-Shock Proteins / genetics
  • HSP70 Heat-Shock Proteins / metabolism
  • HSP90 Heat-Shock Proteins* / genetics
  • HSP90 Heat-Shock Proteins* / metabolism
  • Protein Biosynthesis*
  • Protein Processing, Post-Translational*
  • Proteomics / methods
  • Proteostasis* / genetics
  • Saccharomyces cerevisiae Proteins* / genetics
  • Saccharomyces cerevisiae Proteins* / metabolism
  • Saccharomyces cerevisiae* / genetics
  • Saccharomyces cerevisiae* / metabolism

Substances

  • HSP40 Heat-Shock Proteins
  • Saccharomyces cerevisiae Proteins
  • YDJ1 protein, S cerevisiae
  • HSP90 Heat-Shock Proteins
  • HSP70 Heat-Shock Proteins
  • HSP82 protein, S cerevisiae
  • SSA1 protein, S cerevisiae
  • Adenosine Triphosphatases

Grants and funding

This work was supported by the NIH (R01GM149639 and R01GM139885 to AWT, R35SGM147397 to LF), National Science Foundation Graduate Research Fellowship Program under Grant No. 2235055 (MMM), the Intramural Research Program of the NIH, NCI, Center for Cancer Research (SW). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.