Identification and characterization of Sir2 inhibitors through phenotypic assays in yeast.
Yeast Sir2 is a defining member of a large family of protein deacetylases found in organisms ranging from bacteria to humans. SIR2 was discovered as a gene required for mating in S. cerevisiae 25 years ago, but it was only recently that Sir2's activity as an NAD-dependent protein deacetylase was established. However, years of extensive research did generate a large body of knowledge about the cellular roles of Sir2 in yeast long before its biochemical function was discovered. In addition to Sir2, yeast have four additional NAD-dependent histone deacetylases Hst1-4 (for homologue of Sir2), with distinct cellular roles. Detailed knowledge of the phenotypes of SIR2 and HST loss of function mutants has allowed design of a series of cell based screens that yielded the first inhibitors of NAD-dependent protein deacetylases. These phenotypic assays, amenable to high throughput screening, and coupled with transcript array analysis for evaluation of compound specificity, allowed the identification and detailed characterization of a series of Sir2 inhibitors, entirely bypassing traditional biochemical approaches.
Posakony
Jeff
J
Clinical Research and Human Biology Divisions, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, WA 98109, USA.
Hirao
Maki
M
Bedalov
Antonio
A
D000595:Amino Acid Sequence; D000818:Animals; D020868:Gene Silencing; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009281:Naphthalenes / Q000737:chemistry / Q000494:pharmacology; D010641:Phenotype; D011753:Pyrones / Q000737:chemistry / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D038281:Silent Information Regulator Proteins, Saccharomyces cerevisiae / Q000037:antagonists & inhibitors* / Q000235:genetics*; D056565:Sirtuin 2; D037761:Sirtuins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism*
D056572:Histone Deacetylase Inhibitors; D009281:Naphthalenes; D011753:Pyrones; D029701:Saccharomyces cerevisiae Proteins; D038281:Silent Information Regulator Proteins, Saccharomyces cerevisiae; C444426:splitomicin; C103934:HST1 protein, S cerevisiae; C423033:HST2 protein, S cerevisiae; C087778:SIR2 protein, S cerevisiae; D056565:Sirtuin 2; D037761:Sirtuins; D006655:Histone Deacetylases