Differential recognition of ultraviolet lesions by RecA protein. Possible mechanism for preferential targeting of SOS mutagenesis to (6-4) dipyrimidine sites.
A knowledge of the biochemical basis for UV-induced mutagenesis requires an understanding of the interaction of SOS-activated proteins with DNA polymerase at the replication-blocking dipyrimidine lesions. We have suggested previously that the presence of RecA in this multiprotein complex might be an important feature of induced mutagenesis because RecA associates preferentially with UV-irradiated double-stranded DNA compared to nonirradiated DNA. Previous work by others has indicated that (6-4) dipyrimidine lesions might be more mutagenic than the more common cyclobutane dimer. We have explored the possibility that RecA associates more efficiently with (6-4) lesions than with cyclobutane lesions. We have found that RecA binds DNA with (6-4) lesions much more efficiently than DNA with solely cyclobutane lesions. The distinction between substrates is probably achieved by differential nucleation of the RecA nucleoprotein filament. To investigate the structural basis for differential binding of RecA, we have estimated the unwinding of duplex DNA introduced by (6-4) and cyclobutane lesions. Our data indicate that (6-4) lesions introduce much greater distortion than cyclobutane dimers. We conclude that RecA probably binds preferentially at sites of (6-4) lesions in DNA and that this localization of RecA might target the mutagenic response more frequently to those sites.
Rosenberg
M
M
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Echols
H
H
D010584:Bacteriophage phi X 174 / Q000235:genetics; D004278:DNA, Superhelical / Q000378:metabolism / Q000528:radiation effects*; D004279:DNA, Viral / Q000378:metabolism / Q000528:radiation effects*; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D011740:Pyrimidine Dimers; D011938:Rec A Recombinases / Q000378:metabolism*; D014466:Ultraviolet Rays
D004278:DNA, Superhelical; D004279:DNA, Viral; D011740:Pyrimidine Dimers; D011938:Rec A Recombinases