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PMID: 7784196 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Protein-DNA interactions in soluble telosomes from Saccharomyces cerevisiae.

Nucleic acids research ·Vol. 23 ·No. 9 ·1995-05-11 ·Pages 1454-60

Wright JH, Zakian VA

Abstract

Telomeric DNA in Saccharomyces is organized into a non-nucleosomal chromatin structure called the telosome that can be released from chromosome ends in soluble form by nuclease digestion (Wright, J. H., Gottschling, D. E. and Zakian, V. A. (1992) Genes Dev. 6, 197-210). The protein-DNA interactions of soluble telosomes were investigated by monitoring isolated telomeric DNA fragments for the retention of bound protein using both gel mobility shift and nitrocellulose filter-binding assays. Telosomal proteins remained associated with telomeric DNA at concentrations of ethidium bromide that dissociated nucleosomes. The protein-DNA interactions in the yeast telosome were also disrupted by much lower salt concentrations than those known to disrupt either the interactions of ciliate terminus-binding proteins with telomeric DNA or the interactions of histones with DNA in nucleosomes. Taken together, these data corroborate previously published nuclease mapping data indicating that telosomes are distinct in structure from conventional nucleosomes. These data also indicate that yeast do not possess telomere binding proteins similar to those detected in ciliates that remain tightly bound to telomeric DNA even in high salt. In addition, the characteristic gel mobility shift of soluble telosomes could be mimicked by complexes formed in vitro with yeast telomeric DNA and recombinant Rap1p suggesting that Rap1p, a known component of soluble yeast telosomes (Wright, J. H., Gottschling, D. E. and Zakian, V. A. (1992) Genes Dev. 6, 197-210; Conrad, M. N., Wright, J. H., Wolf, A. J. and Zakian, V. A. (1990) Cell 63, 739-750), is likely to be the major structural protein bound directly to yeast telomeric DNA.

MeSH Terms
DNA, Fungal/metabolism Escherichia coli/genetics Ethidium Fungal Proteins/metabolism GTP-Binding Proteins/genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism Telomere/metabolism rap GTP-Binding Proteins
Chemicals
DNA, Fungal Fungal Proteins GTP-Binding Proteins rap GTP-Binding Proteins Ethidium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wright J H
Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USA.
Zakian V A
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/23.9.1454
Published
1995-05-11
Pages
1454-60
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306882
Subset
IM
Grants
NIGMS NIH HHS · GM43265 · United States
External Links
PubMed source DOI: 10.1093/nar/23.9.1454 PMC Full Text View journal details
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