SOS-dependent replication past a single trans-syn T-T cyclobutane dimer gives a different mutation spectrum and increased error rate compared with replication past this lesion in uninduced cells.
We have transfected SOS-induced and uninduced cells of a uvrA6 strain of Escherichia coli with single-stranded M13mp7-based vectors that carried a single trans-syn T-T cyclobutane dimer at a unique site. Unlike constructs carrying the cis-syn isomer of this lesion, these vectors could be replicated with modest efficiency (14%) in the absence of SOS induction and therefore provided an opportunity to measure directly the influence of such induction on error rate and mutation spectrum. We found that translesion synthesis in the absence of SOS induction was remarkably accurate; only 4% of the replicated bacteriophage contained mutations, which were exclusively targeted single T deletions. In SOS-induced cells, error frequency increased to 11% and the resulting mutations included targeted substitutions and near-targeted single base additions, as well as the T deletions. Replication efficiency was 29% in these conditions. SOS induction therefore leads not only to an enhanced capacity to replicate damaged DNA but also to a marked change in mutation frequency and spectrum.
Banerjee
S K
SK
Department of Biophysics, University of Rochester School of Medicine and Dentistry, New York 14642.
Borden
A
A
Christensen
R B
RB
LeClerc
J E
JE
Lawrence
C W
CW
D001483:Base Sequence; D002872:Chromosome Deletion; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005821:Genetic Techniques; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D011740:Pyrimidine Dimers; D013014:SOS Response, Genetics; D013604:T-Phages / Q000201:enzymology; D014162:Transfection
D015345:Oligonucleotide Probes; D011740:Pyrimidine Dimers; C059224:cyclobutyldithymidine dimers; D004259:DNA-Directed DNA Polymerase