Sequence-specific DNA primer effects on telomerase polymerization activity.
The ribonucleoprotein enzyme telomerase synthesizes one strand of telomeric DNA by copying a template sequence within the RNA moiety of the enzyme. Kinetic studies of this polymerization reaction were used to analyze the mechanism and properties of the telomerase from Tetrahymena thermophila. This enzyme synthesizes TTGGGG repeats, the telomeric DNA sequence of this species, by elongating a DNA primer whose 3' end base pairs with the template-forming domain of the RNA. The enzyme was found to act nonprocessively with short (10- to 12-nucleotide) primers but to become processive as TTGGGG repeats were added. Variation of the 5' sequences of short primers with a common 3' end identified sequence-specific effects which are distinct from those involving base pairing of the 3' end of the primer with the RNA template and which can markedly induce enzyme activity by increasing the catalytic rate of the telomerase polymerization reaction. These results identify an additional mechanistic basis for telomere and DNA end recognition by telomerase in vivo.
Lee
M S
MS
Department of Microbiology and Immunology, University of California, San Francisco 94143-0414.
Blackburn
E H
EH
D000818:Animals; D001483:Base Sequence; D001704:Biopolymers; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D016054:DNA, Protozoan / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid; D013379:Substrate Specificity; D013698:Templates, Genetic; D016808:Tetrahymena thermophila / Q000201:enzymology*
D001704:Biopolymers; D016054:DNA, Protozoan; D004277:DNA, Single-Stranded; D004253:DNA Nucleotidylexotransferase