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A Suppressor Screen for the Characterization of Genetic Links Regulating Chronological Lifespan in Saccharomyces cerevisiae
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Published on: September 17, 2020

A yeast sir2 mutant temperature sensitive for silencing.

Chia-Lin Wang1, Joseph Landry, Rolf Sternglanz

  • 1Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York 11794-5215, USA.

Genetics
|October 11, 2008
PubMed
Summary
This summary is machine-generated.

A mutation in the SIR2 gene causes temperature-sensitive silencing defects in Saccharomyces cerevisiae, impacting mating and gene expression. The mutant Sir2 protein shows reduced deacetylase activity but increased NAD(+) exchange.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Gene silencing is crucial for cellular processes, including mating-type control in Saccharomyces cerevisiae.
  • The SIR2 gene, encoding a histone deacetylase, plays a key role in silencing.
  • Temperature-sensitive mutants are valuable tools for studying essential gene functions.

Purpose of the Study:

  • To identify and characterize new genes involved in silencing in Saccharomyces cerevisiae.
  • To investigate the function of the SIR2 gene and its role in temperature-dependent silencing.
  • To analyze the enzymatic activity of a novel Sir2 mutant protein.

Main Methods:

  • Screening for temperature-sensitive silencing mutants in Saccharomyces cerevisiae.
  • Genetic analysis of a mutant strain with a point mutation in the SIR2 gene.
  • Biochemical assays to compare the enzymatic activity of wild-type and mutant Sir2 proteins.

Main Results:

  • A point mutation (Ser276Cys) in the NAD(+) binding pocket of SIR2 caused temperature-sensitive silencing defects.
  • The mutant exhibited severe mating defects at 37°C but not at 25°C.
  • Mutant Sir2 protein showed reduced deacetylase activity and increased NAD(+)-nicotinamide exchange activity.

Conclusions:

  • The identified SIR2 mutation affects both silencing and enzymatic activity.
  • The NAD(+) binding pocket is critical for Sir2 function, with specific residues influencing deacetylase and exchange activities.
  • This study provides insights into the mechanism of Sir2-mediated gene silencing.