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PMID: 22966204 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Internal amino acids promote Gap1 permease ubiquitylation via TORC1/Npr1/14-3-3-dependent control of the Bul arrestin-like adaptors.

Molecular and cellular biology ·Vol. 32 ·No. 22 ·2012-11-00 ·Pages 4510-22

Merhi A, André B

Abstract

Ubiquitylation of many plasma membrane proteins promotes their endocytosis followed by degradation in the lysosome. The yeast general amino acid permease, Gap1, is ubiquitylated and downregulated when a good nitrogen source like ammonium is provided to cells growing on a poor nitrogen source. This ubiquitylation requires the Rsp5 ubiquitin ligase and the redundant arrestin-like Bul1 and Bul2 adaptors. Previous studies have shown that Gap1 ubiquitylation involves the TORC1 kinase complex, which inhibits the Sit4 phosphatase. This causes inactivation of the protein kinase Npr1, which protects Gap1 against ubiquitylation. However, the mechanisms inducing Gap1 ubiquitylation after Npr1 inactivation remain unknown. We here show that on a poor nitrogen source, the Bul adaptors are phosphorylated in an Npr1-dependent manner and bound to 14-3-3 proteins that protect Gap1 against downregulation. After ammonium is added and converted to amino acids, the Bul proteins are dephosphorylated, dissociate from the 14-3-3 proteins, and undergo ubiquitylation. Furthermore, dephosphorylation of Bul requires the Sit4 phosphatase, which is essential to Gap1 downregulation. The data support the emerging concept that permease ubiquitylation results from activation of the arrestin-like adaptors of the Rsp5 ubiquitin ligase, this coinciding with their dephosphorylation, dissociation from the inhibitory 14-3-3 proteins, and ubiquitylation.

MeSH Terms
14-3-3 Proteins/genetics,metabolism Adaptor Proteins, Signal Transducing/genetics,metabolism Amino Acid Transport Systems/genetics,metabolism Amino Acids/metabolism Gene Expression Regulation, Fungal Lysosomes/metabolism Phosphorylation Protein Kinases/genetics,metabolism Proteolysis Quaternary Ammonium Compounds/metabolism,pharmacology Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Signal Transduction/genetics Transcription Factors/genetics,metabolism Ubiquitin/metabolism Ubiquitin-Protein Ligases/genetics,metabolism Ubiquitination
Chemicals
14-3-3 Proteins Adaptor Proteins, Signal Transducing Amino Acid Transport Systems Amino Acids BUL1 protein, S cerevisiae BUL2 protein, S cerevisiae GAP1 protein, S cerevisiae Quaternary Ammonium Compounds Saccharomyces cerevisiae Proteins TORC1 protein complex, S cerevisiae Transcription Factors Ubiquitin NPR1 protein, S cerevisiae Ubiquitin-Protein Ligases Protein Kinases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Merhi Ahmad
Molecular Physiology of the Cell, Université Libre de Bruxelles, IBMM, Gosselies, Belgium.
André Bruno
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.00463-12
Published
2012-11-00
Epub
2012-00-10
Pages
4510-22
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC3486192
Subset
IM
Corrections
CommentIn
PMID: 23028041
External Links
PubMed source DOI: 10.1128/MCB.00463-12 PMC Full Text View journal details
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