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PMID: 8628672 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

DNA-protein interactions at the telomeric repeats of Schizosaccharomyces pombe.

Nucleic acids research ·Vol. 24 ·No. 8 ·1996-04-15 ·Pages 1412-9

Duffy M, Chambers A

Abstract

Gel retardation assays using a probe containing the repeat region of a Schizosaccharomyces pombe chromosomal telomere identified four specific DNA- protein complexes in S. pombe total protein extracts (I, I', IIa and IIb). The proteins responsible for these complexes bound to the telomeric repeat region irrespective of whether or not the repeats were in close proximity to the end of a DNA molecule, and none of them bound strongly to single-stranded DNA. The protein responsible for complex I (TeRF I) was separated from the activity responsible for complexes IIa and IIb (TeRF II) using heparin-Sepharose chromatography. Both factors were efficiently cross-competed by an oligonucleotide containing the 18 bp sequence 5'-GGTTACAGGTTACAGGTT-3', which corresponds to two complete telomeric repeat units. Mutation of the T residues at positions 4 and 11 in the oligonucleotide dramatically reduced binding by TeRF II, but had no affect on binding by TeRF I. The protein responsible for complex I' did not bind strongly to either the wild-type or mutant oligonucleotide.

MeSH Terms
Base Sequence Binding Sites DNA, Fungal/metabolism DNA, Single-Stranded/metabolism DNA-Binding Proteins/metabolism Fungal Proteins/metabolism Molecular Sequence Data Repetitive Sequences, Nucleic Acid Schizosaccharomyces/genetics,metabolism Telomere
Chemicals
DNA, Fungal DNA, Single-Stranded DNA-Binding Proteins Fungal Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Duffy M
Department of Genetics, University of Nottingham, Queen's Medical Centre, UK.
Chambers A
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/24.8.1412
Published
1996-04-15
Pages
1412-9
Language
English
Region
England
NLM ID
0411011
PMCID
PMC145816
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/24.8.1412 PMC Full Text View journal details
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