Instability of repeated dinucleotides in bacteriophage T7 genomes.
The ligase gene of bacteriophage T7 was interrupted with an insert of synthetic DNA that included a series of dinucleotide repeats which altered the reading frame of the gene and prevented the phage from growing on a host deficient in Escherichia coli ligase. The insert was designed so that gain of an additional copy of the repeat would restore the reading frame and produce ligase positive T7. It was found that a pair of nucleotides was gained at a frequency of 1.4 x 10(-3) and a dinucleotide was lost from this sequence at a frequency of 1 x 10(-4). The same measurements were made using T7 with a DNA polymerase without the 3' --> 5' exonuclease function. This mutant DNA polymerase lacks proofreading edit function and is more processive than its wild-type counterpart. Phage without the edit function gained or lost a dinucleotide repeat significantly more frequently than wild-type T7. Thus, the frequency of two base frameshifting is very high during DNA replication. Proofreading activity attenuates the frequency of two base frameshifts. Inactivation of the 3' --> 5' exonuclease of T7 DNA polymerase had essentially no effect on the frequency of deletion of one member of a pair of 10 bp tandem repeats.
Yang
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Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Masker
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W
D017123:Bacteriophage T7 / Q000235:genetics*; D001483:Base Sequence; D004261:DNA Replication; D018900:Dinucleotide Repeats; D016679:Genome, Viral; D008969:Molecular Sequence Data; D009154:Mutation