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PMID: 3865186 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Genetic complementation of UV-induced DNA repair in Chinese hamster ovary cells by the denV gene of phage T4.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 22 ·1985-11-00 ·Pages 7656-60

Valerie K, de Riel JK, Henderson EE

Abstract

The denV gene of phage T4, encoding the pyrimidine dimer-specific DNA repair enzyme endonuclease V, has been introduced by DNA transfection into the UV-sensitive DNA repair-deficient Chinese hamster ovary (CHO) cell line UV5. Transformants were first selected for resistance to the antibiotic G418 conferred by the neo gene from Tn5 carried by the same plasmid. A majority of the isolated G418-resistant UV5 clones also showed an increased resistance to 254-nm UV light. One clone, designated I-A1, was found to have an intermediate level of colony-forming ability after UV irradiation when compared to UV5 and wild-type AA8 cells. A Southern blot showed that I-A1 carries a single integrated intact copy of the denV gene. Alkaline sucrose gradients revealed a dose-dependent appearance of breaks in the DNA of I-A1 cells following UV-irradiation, while unirradiated cells did not exhibit any significant breaks. Analysis of DNA repair by isopycnic sedimentation showed that DNA excision repair by I-A1 was at least equal to the level of repair in AA8 cells. These results show that the prokaryotic denV gene can restore UV repair capabilities in vivo to CHO UV5 cells defective in repair of UV-induced damage.

MeSH Terms
Animals Cell Survival/radiation effects Cells, Cultured Cricetinae Cricetulus DNA/biosynthesis,radiation effects DNA Repair Deoxyribonuclease (Pyrimidine Dimer) Endodeoxyribonucleases/genetics Female Genes, Viral Genetic Complementation Test Genetic Vectors Ovary T-Phages/genetics Transfection Ultraviolet Rays
Chemicals
DNA Endodeoxyribonucleases Deoxyribonuclease (Pyrimidine Dimer)
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Valerie K
de Riel J K
Henderson E E
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31 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.82.22.7656
Published
1985-11-00
Pages
7656-60
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC391392
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.82.22.7656 PMC Full Text View journal details
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