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PMID: 15060140 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vivo dynamics of Swi6 in yeast: evidence for a stochastic model of heterochromatin.

Molecular and cellular biology ·Vol. 24 ·No. 8 ·2004-04-00 ·Pages 3157-67

Cheutin T, Gorski SA, May KM, Singh PB, Misteli T

Abstract

The mechanism for transcriptional silencing of pericentric heterochromatin is conserved from fission yeast to mammals. Silenced genome regions are marked by epigenetic methylation of histone H3, which serves as a binding site for structural heterochromatin proteins. In the fission yeast Schizosaccharomyces pombe, the major structural heterochromatin protein is Swi6. To gain insight into Swi6 function in vivo, we have studied its dynamics in the nucleus of living yeast. We demonstrate that, in contrast to mammalian cells, yeast heterochromatin domains undergo rapid, large-scale motions within the nucleus. Similar to the situation in mammalian cells, Swi6 does not permanently associate with these chromatin domains but binds only transiently to euchromatin and heterochromatin. Swi6 binding dynamics are dependent on growth status and on the silencing factors Clr4 and Rik1, but not Clr1, Clr2, or Clr3. By comparing the kinetics of mutant Swi6 proteins in swi6(-) and swi6(+) strains, we demonstrate that homotypic protein-protein interactions via the chromoshadow domain stabilize Swi6 binding to chromatin in vivo. Kinetic modeling allowed quantitative estimation of residence times and indicated the existence of at least two kinetically distinct populations of Swi6 in heterochromatin. The observed dynamics of Swi6 binding are consistent with a stochastic model of heterochromatin and indicate evolutionary conservation of heterochromatin protein binding properties from mammals to yeast.

MeSH Terms
Animals Chromosomal Proteins, Non-Histone/genetics,metabolism Evolution, Molecular Fluorescence Recovery After Photobleaching Gene Silencing Heterochromatin Models, Genetic Nuclear Proteins/genetics,metabolism Nucleosomes/metabolism Protein Binding Recombinant Fusion Proteins/genetics,metabolism Schizosaccharomyces/genetics,metabolism Schizosaccharomyces pombe Proteins/genetics,metabolism
Chemicals
Chromosomal Proteins, Non-Histone Heterochromatin Nuclear Proteins Nucleosomes Recombinant Fusion Proteins Schizosaccharomyces pombe Proteins Swi6 protein, S pombe
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Cheutin Thierry
National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Gorski Stanislaw A
May Karen M
Singh Prim B
Misteli Tom
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.24.8.3157-3167.2004
Published
2004-04-00
Pages
3157-67
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC381678
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.24.8.3157-3167.2004 PMC Full Text View journal details
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