UV-enhanced expression of a reporter gene is induced at lower UV fluences in transcription-coupled repair deficient compared to normal human fibroblasts, and is absent in SV40-transformed counterparts.
UV irradiation enhances transcription of a number of cellular and viral genes. We have compared dose responses for alterations in expression from reporter constructs driven by the human and murine cytomegalovirus (CMV) immediate early (IE) promoters in cells from patients with deficiencies in nucleotide excision repair (complementation groups of xeroderma pigmentosum and Cockayne syndrome) following UV exposure, or infection with UV-damaged recombinant vectors. Results suggest that unrepaired damage in active genes triggers increased reporter activity from constructs driven by the CMV promoters in human fibroblasts. Similar to human fibroblasts, HeLa cells and cells from Li-Fraumeni syndrome patients (characterized by an inherited mutation in the p53 gene) also displayed an increase in reporter activity following UV exposure; however, this response was absent in all simian virus 40 (SV40)-transformed cell lines examined. This suggests that a pathway affected by SV40-transformation (other than p53) plays an essential role in UV-enhanced expression from the CMV IE promoter.
Francis
M A
MA
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
Rainbow
A J
AJ
D000818:Animals; D002460:Cell Line; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral / Q000235:genetics*; D003587:Cytomegalovirus / Q000235:genetics; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000378:metabolism / Q000528:radiation effects* / Q000821:virology; D015870:Gene Expression / Q000528:radiation effects; D017930:Genes, Reporter / Q000528:radiation effects*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D013539:Simian virus 40 / Q000235:genetics*; D014158:Transcription, Genetic / Q000528:radiation effects; D014466:Ultraviolet Rays