Sequencing of Saccharomyces telomeres cloned using T4 DNA polymerase reveals two domains.
By using T4 DNA polymerase rather than S1 or Bal31 nuclease to clone yeast telomeres, very little telomeric DNA is lost. These clones were used to determine the DNA sequence of virtually the entire telomeric tract. Our results demonstrated that a slightly modified version, C2-3A(CA)1-6, of the consensus derived from sequence analysis of more-internal regions (J. Shampay, J. W. Szostak, and E. H. Blackburn, Nature [London] 310:154-157, 1984) extends to the very end of the chromosome. The sequence analysis also suggests that yeast telomeres consist of two domains: the proximal 120 to 150 base pairs, which appear to be protected from processes such as recombination, degradation, and elongation, and the distal portion of the telomere, which is more susceptible to these events.
Wang
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SS
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Zakian
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VA
D001483:Base Sequence; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular / Q000379:methods*; D004271:DNA, Fungal / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013604:T-Phages / Q000201:enzymology
D004271:DNA, Fungal; D004259:DNA-Directed DNA Polymerase