Yeast α-arrestin Art2 is the key regulator of ubiquitylation-dependent endocytosis of plasma membrane vitamin B1 transporters

Jérôme Savocco, Sylvain Nootens, Wilhelmine Afokpa, Mathilde Bausart, Xiaoqian Chen, Jennifer Villers, Henri François Renard, Martine Prévost, Ruddy Wattiez, Pierre Morsomme

Research output: Contribution to journalArticle

Abstract

Endocytosis of membrane proteins in yeast requires α-arrestin-mediated ubiquitylation by the ubiquitin ligase Rsp5. Yet, the diversity of α-arrestin targets studied is restricted to a small subset of plasma membrane (PM) proteins. Here, we performed quantitative proteomics to identify new targets of 12 α-arrestins and gained insight into the diversity of pathways affected by α-arrestins, including the cell wall integrity pathway and PM–endoplasmic reticulum contact sites. We found that Art2 is the main regulator of substrate- and stress-induced ubiquitylation and endocytosis of the thiamine (vitamin B1) transporters: Thi7, nicotinamide riboside transporter 1 (Nrt1), and Thi72. Genetic screening allowed for the isolation of transport-defective Thi7 mutants, which impaired thiamine-induced endocytosis. Coexpression of inactive mutants with wild-type Thi7 revealed that both transporter conformation and transport activity are important to induce endocytosis. Finally, we provide evidence that Art2 mediated Thi7 endocytosis is regulated by the target of rapamycin complex 1 (TORC1) and requires the Sit4 phosphatase but is not inhibited by the Npr1 kinase.

Original languageEnglish
Article numbere3000512
Pages (from-to)e3000512
JournalPLoS biology
Volume17
Issue number10
DOIs
Publication statusPublished - 1 Oct 2019
Externally publishedYes

Keywords

  • Arrestins/genetics
  • Cell Membrane/drug effects
  • Cell Wall/drug effects
  • Endocytosis/genetics
  • Endoplasmic Reticulum/drug effects
  • Endosomal Sorting Complexes Required for Transport/genetics
  • Gene Expression Regulation, Fungal
  • Membrane Transport Proteins/genetics
  • Models, Molecular
  • Mutation
  • Nucleoside Transport Proteins/genetics
  • Protein Binding
  • Protein Kinases/genetics
  • Protein Phosphatase 2/genetics
  • Protein Processing, Post-Translational
  • Protein Structure, Secondary
  • Proteomics/methods
  • Saccharomyces cerevisiae/drug effects
  • Saccharomyces cerevisiae Proteins/genetics
  • Signal Transduction
  • Thiamine/metabolism
  • Transcription Factors/genetics
  • Ubiquitin-Protein Ligase Complexes/genetics
  • Ubiquitination

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  • Cite this

    Savocco, J., Nootens, S., Afokpa, W., Bausart, M., Chen, X., Villers, J., Renard, H. F., Prévost, M., Wattiez, R., & Morsomme, P. (2019). Yeast α-arrestin Art2 is the key regulator of ubiquitylation-dependent endocytosis of plasma membrane vitamin B1 transporters. PLoS biology, 17(10), e3000512. [e3000512]. https://doi.org/10.1371/journal.pbio.3000512