Telomerase is the ribonucleoprotein reverse transcriptase that adds telomeric DNA repeats to the ends of chromosomes. This involves annealing of the telomerase RNA template to the 30 end of the chromosome, reverse transcription of the RNA template by the telomerase reverse transcriptase polypeptide and translocation. Here, we overexpress and partially purify the catalytically active yeast telomerase core in its natural host and probe telomerase RNA base methylation accessibility with dimethyl sulphate in the presence and absence of a DNA substrate and after substrate elongation. The length of the RNA - DNA hybrid is kept constant at seven base pairs after primer binding and elongation. Thus, new base-pair formation at the 30 end of the substrate during elongation coincides with disruption of base-pair interactions at the other side of the template. Presumably, this circumvents the generation of an exceedingly high energy barrier for translocation and dissociation. Our analysis also corroborates recently proposed yeast telomerase RNA secondary structure models.
机构:
UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
BHATTACHARYYA, A
;
BLACKBURN, EH
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UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
Bryan, TM
;
Goodrich, KJ
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机构:
Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
Goodrich, KJ
;
Cech, TR
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机构:
Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
机构:
UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
COHN, M
;
BLACKBURN, EH
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h-index: 0
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UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USA
Hammond, PW
;
Cech, TR
论文数: 0引用数: 0
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机构:
Univ Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USA
机构:
UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
BHATTACHARYYA, A
;
BLACKBURN, EH
论文数: 0引用数: 0
h-index: 0
机构:
UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
Bryan, TM
;
Goodrich, KJ
论文数: 0引用数: 0
h-index: 0
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
Goodrich, KJ
;
Cech, TR
论文数: 0引用数: 0
h-index: 0
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Chem & Biochem, Boulder, CO 80309 USA
机构:
UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
COHN, M
;
BLACKBURN, EH
论文数: 0引用数: 0
h-index: 0
机构:
UNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USAUNIV CALIF SAN FRANCISCO, DEPT MICROBIOL & IMMUNOL, SAN FRANCISCO, CA 94143 USA
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USA
Hammond, PW
;
Cech, TR
论文数: 0引用数: 0
h-index: 0
机构:
Univ Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USAUniv Colorado, Howard Hughes Med Inst, Dept Biochem & Chem, Boulder, CO 80309 USA