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PMID: 18362167 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

The silent information regulator 3 protein, SIR3p, binds to chromatin fibers and assembles a hypercondensed chromatin architecture in the presence of salt.

Molecular and cellular biology ·Vol. 28 ·No. 11 ·2008-06-00 ·Pages 3563-72

McBryant SJ, Krause C, Woodcock CL, Hansen JC

Abstract

The telomeres and mating-type loci of budding yeast adopt a condensed, heterochromatin-like state through recruitment of the silent information regulator (SIR) proteins SIR2p, SIR3p, and SIR4p. In this study we characterize the chromatin binding determinants of recombinant SIR3p and identify how SIR3p mediates chromatin fiber condensation in vitro. Purified full-length SIR3p was incubated with naked DNA, nucleosome core particles, or defined nucleosomal arrays, and the resulting complexes were analyzed by electrophoretic shift assays, sedimentation velocity, and electron microscopy. SIR3p bound avidly to all three types of templates. SIR3p loading onto its nucleosomal sites in chromatin produced thickened condensed fibers that retained a beaded morphology. At higher SIR3p concentrations, individual nucleosomal arrays formed oligomeric suprastructures bridged by SIR3p oligomers. When condensed SIR3p-bound chromatin fibers were incubated in Mg(2+), they folded and oligomerized even further to produce hypercondensed higher-order chromatin structures. Collectively, these results define how SIR3p may function as a chromatin architectural protein and provide new insight into the interplay between endogenous and protein-mediated chromatin fiber condensation pathways.

MeSH Terms
Chromatin/chemistry,ultrastructure DNA/chemistry,ultrastructure Nucleosomes/chemistry Recombinant Proteins/chemistry,genetics Salts/chemistry Silent Information Regulator Proteins, Saccharomyces cerevisiae/chemistry,genetics
Chemicals
Chromatin Nucleosomes Recombinant Proteins SIR3 protein, S cerevisiae Salts Silent Information Regulator Proteins, Saccharomyces cerevisiae DNA
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
McBryant Steven J
Department of Biochemistry and Molecular Biology, Colorado State University, 381 MRB, Fort Collins, CO 80523-1870, USA. smcbryan@lamar.colostate.edu
Krause Christine
Woodcock Christopher L
Hansen Jeffrey C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01389-07
Email
smcbryan@lamar.colostate.edu
Published
2008-06-00
Epub
2008-00-24
Pages
3563-72
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2423291
Subset
IM
Grants
NIGMS NIH HHS · R01 GM066834 · United States
NIGMS NIH HHS · GM43786 · United States
NIGMS NIH HHS · GM66834 · United States
External Links
PubMed source DOI: 10.1128/MCB.01389-07 PMC Full Text View journal details
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