Identification and characterization of a bovine sperm protein that binds specifically to single-stranded telomeric deoxyribonucleic acid.
Telomere DNA at the physical termini of chromosomes forms a single-stranded 3' overhang. In lower eukaryotes, e.g., ciliated protozoa, this DNA extension is capped by specific proteins that have been structurally and functionally characterized. Much less is known about single-stranded telomere DNA-binding proteins in vertebrates. Here we describe a new protein from bovine sperm designated bsSSTBP that specifically interacts with single-stranded (TTAGGG)(N) DNA. The bsSSTBP was extracted from nuclei by 0.6 M KCl. The native size of this protein, estimated by gel filtration, was 20-40 kDa. SDS-PAGE of the UV cross-linked complex between bsSSTBP and telomere DNA indicated that several polypeptides are involved in complex formation. Bovine sSSTB had high specificity toward nucleotide sequence, since single nucleotide substitutions in the (TTAGGG)(4) substrate suppressed binding. The minimal number of (TTAGGG) repeats required for binding of bsSSTBP was 3, and the protein recognized linear but not folded DNA structures. We propose that the bsSSTBP participates in telomere-telomere interactions and the telomere membrane localization observed in mature sperm. In mammals, somatic telomere-binding proteins are apparently substituted by sperm-specific ones that may lead to a structural reorganization of telomere domains to fulfill functions important during meiosis and fertilization.
Kozik
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Department of Biological Chemistry, School of Medicine, University of California at Davis, Davis, California 95616, USA.
Bradbury
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EM
Zalensky
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AO
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D008297:Male; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D013094:Spermatozoa / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D016615:Telomere / Q000378:metabolism* / Q000648:ultrastructure; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded; D015345:Oligonucleotide Probes; D011506:Proteins