Restoration of u.v.-induced excision repair in Xeroderma D cells transfected with the denV gene of bacteriophage T4.
The heritable DNA repair defect in human Xeroderma D cells, which results in failure to incise at u.v. light-induced pyrimidine dimers, has been partially but stably corrected by transfection of immortalised cells with the denV pyrimidine dimer glycosylase gene of bacteriophage T4. Transfectants selected either for a dominant marker on the mammalian vector carrying the prokaryotic gene or for the dominant marker plus resistance to killing by u.v. light, have been shown to express the denV gene to varying degrees. denV expression results in significant phenotypic change in the initially repair-deficient, u.v.-hypersensitive cells. Increased resistance to u.v. light and more rapid recovery of replicative DNA synthesis following u.v. irradiation have been correlated both with improved repair DNA synthesis and with a novel dimer incision capability present in denV transfected Xeroderma cells but not as evident in transfected normal cells. Most of the transfectants contain a single integrated copy of the denV gene; increase in denV copy number does not result in either increased gene expression or enhanced survival to u.v. light. These results show that expression of a heterologous prokaryotic repair gene can partially compensate for the genetic defect in a human Xeroderma D cell.
Arrand
J E
JE
Department of Zoology, University of Cambridge, UK.
Squires
S
S
Bone
N M
NM
Johnson
R T
RT
D002460:Cell Line; D045647:DNA Glycosylases; D004260:DNA Repair / Q000528:radiation effects*; D004261:DNA Replication / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005814:Genes, Viral; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism; D007700:Kinetics; D009699:N-Glycosyl Hydrolases / Q000235:genetics*; D013604:T-Phages / Q000235:genetics*; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; C036401:deoxyribopyrimidine endonucleosidase