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PMID: 9122169 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Silent information regulator protein complexes in Saccharomyces cerevisiae: a SIR2/SIR4 complex and evidence for a regulatory domain in SIR4 that inhibits its interaction with SIR3.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 94 ·No. 6 ·1997-03-18 ·Pages 2186-91

Moazed D, Kistler A, Axelrod A, Rine J, Johnson AD

Abstract

The SIR2, SIR3, and SIR4 silent information regulator proteins are involved in the assembly of silent chromatin domains in the budding yeast Saccharomyces cerevisiae. Using a series of biochemical experiments, we have studied protein-protein interactions involving these proteins. We found that yeast extracts contained a SIR2/SIR4 complex that was associated with little or no SIR3. However, truncations of the N-terminal two-thirds of the SIR4 protein allowed it to efficiently associate with SIR3, suggesting that the N-terminal domain of SIR4 inhibited its interaction with SIR3. We propose that the SIR3 and SIR4 proteins interact only during the assembly of the SIR protein complex at the silencer and that an early step in assembly unmasks the SIR4 protein to allow its association with SIR3. To test whether the interactions observed in yeast extracts were direct, we tested these SIR-SIR interactions using bacterially expressed SIR proteins. We observed direct interactions between SIR4 and SIR2, SIR4 and SIR3, SIR2 and SIR3, SIR2 and SIR2, and SIR4 and SIR4, indicating that the associations observed in yeast extracts were direct.

MeSH Terms
Binding Sites Cloning, Molecular DNA-Binding Proteins/chemistry,isolation & purification,metabolism Escherichia coli Fungal Proteins/chemistry,isolation & purification,metabolism Glutathione Transferase Histone Deacetylases Models, Structural Recombinant Fusion Proteins/chemistry,isolation & purification,metabolism Restriction Mapping Saccharomyces cerevisiae/genetics,metabolism Silent Information Regulator Proteins, Saccharomyces cerevisiae Sirtuin 2 Sirtuins Trans-Activators/chemistry,isolation & purification,metabolism
Chemicals
DNA-Binding Proteins Fungal Proteins Recombinant Fusion Proteins SIR3 protein, S cerevisiae SIR4 protein, S cerevisiae Silent Information Regulator Proteins, Saccharomyces cerevisiae Trans-Activators Glutathione Transferase SIR2 protein, S cerevisiae Sirtuin 2 Sirtuins Histone Deacetylases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Moazed D
Department of Biochemistry and Biophysics, University of California, San Francisco 94143, USA.
Kistler A
Axelrod A
Rine J
Johnson A D
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.94.6.2186
Published
1997-03-18
Pages
2186-91
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC20062
Subset
IM
Grants
NIGMS NIH HHS · R37 GM031105 · United States
NIGMS NIH HHS · GM53452-01 · United States
NIGMS NIH HHS · R01 GM031105 · United States
NCI NIH HHS · T32 CA009270 · United States
NCI NIH HHS · T32 CA09270 · United States
NIGMS NIH HHS · GM31105 · United States
External Links
PubMed source DOI: 10.1073/pnas.94.6.2186 PMC Full Text View journal details
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