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PMID: 11500379 Published · ppublish English Journal Article

Sir2p exists in two nucleosome-binding complexes with distinct deacetylase activities.

The EMBO journal ·Vol. 20 ·No. 16 ·2001-08-15 ·Pages 4522-35

Ghidelli S, Donze D, Dhillon N, Kamakaka RT

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,isolation & purification,metabolism,physiology Histone Deacetylases/genetics,isolation & purification,metabolism,physiology Mutagenesis Nuclear Proteins/metabolism Nucleosomes/metabolism Recombinant Fusion Proteins/genetics,isolation & purification,metabolism,physiology Saccharomyces cerevisiae Proteins Silent Information Regulator Proteins, Saccharomyces cerevisiae Sirtuin 2 Sirtuins Trans-Activators/genetics,isolation & purification,metabolism,physiology
Chemicals
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
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ghidelli S
Unit on Chromatin and Transcription, NICHD/NIH, Building 18T, Room 106, 18 Library Drive, Bethesda, MD 20892, USA.
Donze D
Dhillon N
Kamakaka R T
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.16.4522
Published
2001-08-15
Pages
4522-35
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125569
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.16.4522 PMC Full Text View journal details
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