Mutational analyses of dinucleotide and tetranucleotide microsatellites in Escherichia coli: influence of sequence on expansion mutagenesis.
Mutagenesis at [GT/CA](10), [TC/AG](11) and [TTCC/AAGG](9) microsatellite sequences inserted in the herpes simplex virus thymidine kinase (HSV-tk) gene was analyzed in isogenic mutL(+) and mutL(-) Escherichia coli. In both strains, significantly more expansion than deletion mutations were observed at the [TTCC/AAGG](9) motif relative to either dinucleotide motif. As the HSV-tk coding sequence contains an endogenous [G/C](7) mononucleotide repeat and approximately 1000 bp of unique sequence, we were able to compare mutagenesis among various sequence motifs. We observed that the relative risk of mutation in E.COLI: is: [TTCC/AAGG](9) > [GT/CA](10) approximately [TC/AG](11) > unique approximately [G/C](7). The mutation frequency varied 1400-fold in mutL(+) cells between the tetranucleotide motif and the mononucleotide motif, but only 50-fold in mutL(-) cells. The [G/C](7) sequence was destabilized the greatest and the tetranucleotide motif the least by loss of mismatch repair. These results demonstrate that the quantitative risk of mutation at various microsatellites greatly depends on the DNA sequence composition. We suggest alternative models for the production of expansion mutations during lagging strand replication of the [TTCC/AAGG](9) microsatellite.
Eckert
K A
KA
The Jake Gittlen Cancer Research Institute, The Pennsylvania State University College of Medicine, PO Box 850, Hershey, PA 17033, USA. kae4@psu.edu
Yan
G
G
D000251:Adenosine Triphosphatases; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D020137:Base Pair Mismatch; D004260:DNA Repair; D004274:DNA, Recombinant / Q000235:genetics; D018900:Dinucleotide Repeats / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D018895:Microsatellite Repeats / Q000235:genetics*; D000070956:MutL Proteins; D009154:Mutation; D010957:Plasmids; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000235:genetics
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D029968:Escherichia coli Proteins; C074770:MutL protein, E coli; D013937:Thymidine Kinase; D000251:Adenosine Triphosphatases; D000070956:MutL Proteins