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PMID: 15071494 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The absence of the yeast chromatin assembly factor Asf1 increases genomic instability and sister chromatid exchange.

EMBO reports ·Vol. 5 ·No. 5 ·2004-05-00 ·Pages 497-502

Prado F, Cortés-Ledesma F, Aguilera A

Abstract

Histone chaperone Asf1 participates in heterochromatin silencing, DNA repair and regulation of gene expression, and promotes the assembly of DNA into chromatin in vitro. To determine the influence of Asf1 on genetic stability, we have analysed the effect of asf1Delta on homologous recombination. In accordance with a defect in nucleosome assembly, asf1Delta leads to a loss of negative supercoiling in plasmids. Importantly, asf1Delta increases spontaneous recombination between inverted DNA sequences. This increase correlates with an accumulation of double-strand breaks (DSBs) as determined by immunodetection of phosphorylated histone H2A and fluorescent detection of Rad52-YFP foci during S and G2/M phases. In addition, asf1Delta shows high levels of sister chromatid exchange (SCE) and is proficient in DSB-induced SCE as determined by physical analysis. Our results suggest that defective chromatin assembly caused by asf1Delta leads to DSBs that can be repaired by SCE, affecting genetic stability.

MeSH Terms
Cell Cycle Proteins/genetics,metabolism Chromatin/metabolism DNA Damage DNA Repair DNA-Binding Proteins/genetics,metabolism Genomic Instability Molecular Chaperones/genetics,metabolism Nucleic Acid Conformation Nucleosomes/metabolism Plasmids/genetics Rad52 DNA Repair and Recombination Protein Recombinant Fusion Proteins/genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Sister Chromatid Exchange
Chemicals
ASF1 protein, S cerevisiae Cell Cycle Proteins Chromatin DNA-Binding Proteins Molecular Chaperones Nucleosomes RAD52 protein, S cerevisiae Rad52 DNA Repair and Recombination Protein Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Prado Félix
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Avda. Reina Mercedes 6, 41012 Sevilla, Spain.
Cortés-Ledesma Felipe
Aguilera Andrés
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Article Info
Journal
EMBO reports
Abbr.
EMBO Rep
ISSN
1469-221X
DOI
10.1038/sj.embor.7400128
Published
2004-05-00
Epub
2004-00-08
Pages
497-502
Language
English
Region
England
NLM ID
100963049
PMCID
PMC1299049
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.embor.7400128 PMC Full Text View journal details
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