Incomplete complementation of the DNA repair defect in cockayne syndrome cells by the denV gene from bacteriophage T4 suggests a deficiency in base excision repair.
Endonuclease V (denV) from bacteriophage T4 has been examined for its ability to complement the repair defect in Cockayne syndrome (CS) cells of complementation groups A and B. CS is an autosomal recessive disorder characterized by hypersensitivity to UV light and a defect in the preferential repair of UV-induced lesions in transcriptionally active DNA by the nucleotide excision repair (NER) pathway. The denV gene was introduced into non-transformed normal and CS fibroblasts transiently via a recombinant adenovirus (Ad) vector and into SV40-transformed normal and CS cells via a retroviral vector. Expression of denV in CS-A cells resulted in partial correction of the UV-sensitive phenotype in assays of gene-specific repair and cell viability, while correction of CS-B cells by expression of denV in the same assays was minimal or non-existent. In contrast, denV expression led to enhanced host cell reactivation (HCR) of viral DNA synthesis in both CS complementation groups to near normal levels. DenV is a glycosylase which is specific for cyclobutane-pyrimidine dimers (CPDs) but does not recognize other UV-induced lesions. Previous work has indicated that CS cells can efficiently repair all non-CPD UV-induced transcription blocking lesions (S.F. Barrett et al.. Mutation Res. 255 (1991) 281-291 [1]) and that denV incised lesions are believed to be processed via the base excision repair (BER) pathway. The inability of denV to complement the NER defect in CS cells to normal levels implies an impaired ability to process denV incised lesions by the BER pathway, and suggests a role for the CS genes, particularly the CS-B gene, in BER.
Francis
M A
MA
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
Bagga
P S
PS
Athwal
R S
RS
Rainbow
A J
AJ
D000260:Adenoviruses, Human / Q000235:genetics; D017122:Bacteriophage T4 / Q000201:enzymology; D002460:Cell Line; D002461:Cell Line, Transformed; D003057:Cockayne Syndrome / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics*; D004279:DNA, Viral / Q000096:biosynthesis; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004706:Endodeoxyribonucleases / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D005347:Fibroblasts / Q000528:radiation effects; D018014:Gene Transfer Techniques; D005814:Genes, Viral / Q000235:genetics / Q000502:physiology; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006801:Humans; D012333:RNA, Messenger / Q000032:analysis; D012190:Retroviridae / Q000235:genetics; D013539:Simian virus 40; D012867:Skin / Q000166:cytology; D014466:Ultraviolet Rays; D015678:Viral Structural Proteins / Q000235:genetics
D004279:DNA, Viral; D012333:RNA, Messenger; D015678:Viral Structural Proteins; D004706:Endodeoxyribonucleases; D043224:Deoxyribonuclease (Pyrimidine Dimer)