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The deubiquitinating enzyme Dot4p is involved in regulating nutrient uptake
1Pritzker School of Medicine, University of Chicago, 924 East 57th Street, Chicago, Illinois, USA. kah7@midway.uchicago.edu
Biochemical and Biophysical Research Communications
|May 16, 2001
Summary
DOT4 gene mutations in yeast disrupt nutrient transport regulation. Preventing endocytosis partially rescued growth defects, indicating Dot4p
Area of Science:
- Molecular biology
- Cell biology
- Yeast genetics
Background:
- Ubiquitination regulates yeast nutrient transporters for internalization and degradation.
- DOT4 gene encodes a deubiquitinating enzyme crucial for protein homeostasis.
Purpose of the Study:
- Investigate the role of DOT4 in nutrient transporter regulation in yeast.
- Determine the mechanism by which DOT4 mutations affect cell growth and nutrient transport.
Main Methods:
- Genetic analysis of DOT4 mutants in yeast.
- Assessing cell growth defects under nutrient limitation.
- Monitoring endocytosis of membrane proteins.
- Measuring amino acid permease Gap1p activity and protein levels.
Main Results:
- Mutations in DOT4 cause severe growth defects, especially when combined with nutrient biosynthetic defects.
- Inhibiting endocytosis partially suppressed the dot4Δ growth defect.
- DOT4 mutants exhibit reduced amino acid permease Gap1p activity and protein levels.
- GAP1 mRNA levels remain unchanged in DOT4 mutants, suggesting posttranscriptional regulation.
Conclusions:
- Dot4p plays a critical role in the posttranscriptional regulation of the amino acid permease Gap1p.
- Dot4p likely influences nutrient transporter stability and function through deubiquitination.
- These findings highlight the importance of deubiquitinating enzymes in maintaining cellular nutrient homeostasis.