Abstract
The absolute requirement for the histone deacetylase activity of Sir2p in silencing coupled with the conservation of Sir2p-like proteins in larger eukaryotes suggests that this molecule plays an important role in gene regulation in all organisms. Here we report the purification and characterization of two Sir2p-containing protein complexes; one of which contains Sir4p and the other Net1p. The Sir4p-containing complex has an NAD-dependent histone deacetylase activity, while the Net1p-containing complex possesses deacetylase activity but only weak NAD-dependent histone deacetylase activity. Finally, we demonstrate that the Sir2p-containing complexes bind nucleosomes efficiently and partially restrict accessibility of the linker DNA to enzymatic probes.
MeSH terms
-
Binding Sites
-
Cell Cycle Proteins*
-
Fungal Proteins / genetics
-
Fungal Proteins / isolation & purification
-
Fungal Proteins / metabolism
-
Fungal Proteins / physiology*
-
Histone Deacetylases / genetics
-
Histone Deacetylases / isolation & purification
-
Histone Deacetylases / metabolism
-
Histone Deacetylases / physiology*
-
Mutagenesis
-
Nuclear Proteins / metabolism
-
Nucleosomes / metabolism*
-
Recombinant Fusion Proteins / genetics
-
Recombinant Fusion Proteins / isolation & purification
-
Recombinant Fusion Proteins / metabolism
-
Recombinant Fusion Proteins / physiology
-
Saccharomyces cerevisiae Proteins*
-
Silent Information Regulator Proteins, Saccharomyces cerevisiae*
-
Sirtuin 2
-
Sirtuins
-
Trans-Activators / genetics
-
Trans-Activators / isolation & purification
-
Trans-Activators / metabolism
-
Trans-Activators / physiology*
Substances
-
Cell Cycle Proteins
-
Fungal Proteins
-
Net1 protein, S cerevisiae
-
Nuclear Proteins
-
Nucleosomes
-
Recombinant Fusion Proteins
-
SIR3 protein, S cerevisiae
-
SIR4 protein, S cerevisiae
-
Saccharomyces cerevisiae Proteins
-
Silent Information Regulator Proteins, Saccharomyces cerevisiae
-
Trans-Activators
-
SIR2 protein, S cerevisiae
-
Sirtuin 2
-
Sirtuins
-
Histone Deacetylases