Preirradiation of host cells does not alter blockage of simian virus 40 replication forks by pyrimidine dimers.
Do damage-inducible responses in mammalian cells alter the interaction of lesions with replication forks? We have previously demonstrated that preirradiation of the host cell mitigates UV inhibition of SV40 DNA replication; this mitigation can be detected within the first 30 min after the test irradiation. Here we test the hypotheses that this mitigation involves either (1) rapid dimer removal, (2) rapid synthesis of daughter strands past lesions (trans-dimer synthesis), or (3) continued progression of the replication fork beyond a dimer. Cells preirradiated with UV were infected with undamaged SV40, and the effects of UV upon viral DNA synthesis were measured within the first hour after a subsequent test irradiation. In preirradiated cells, as well as in non-preirradiated cells, pyrimidine dimers block elongation of daughter strands; daughter strands grow only to a size equal to the interdimer distance along the parental strands. There is, within this first hour after UV, no evidence for trans-dimer synthesis, nor for more rapid dimer removal either in the bulk of the parental DNA or in molecules in the replication pool. Progression of the replication forks was analyzed by electron microscopy of replicating SV40 molecules. Dimers block replication-fork progression in preirradiated cells to the same extent as in non-preirradiated cells. These experiments argue strongly against the hypotheses that preirradiation of host cells results in either the rapid removal of dimers, trans-dimer synthesis, or continued replication-fork progression beyond dimers.
Scaria
A
A
Department of Biochemistry, Indiana University School of Medicine, Indianapolis 46223.
Edenberg
H J
HJ
D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects*; D004279:DNA, Viral / Q000096:biosynthesis; D008854:Microscopy, Electron; D011740:Pyrimidine Dimers / Q000502:physiology*; D013539:Simian virus 40 / Q000235:genetics / Q000528:radiation effects*; D013997:Time Factors; D014466:Ultraviolet Rays; D014779:Virus Replication / Q000528:radiation effects*
D004279:DNA, Viral; D011740:Pyrimidine Dimers