Molecular Cloning and Characterization of a Mammalian Excision Repair Gene that Partially Restores UV Resistance to Xeroderma Pigmentosum Complementation Group D Cells

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 86; no. 18; pp. 6997 - 7001
• Main Authors: Arrand, Janet E., Bone, Neil M., Johnson, Robert T.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-09-1989; National Acad Sciences
• Subjects: 560120 - Radiation Effects on Biochemicals, Cells, & Tissue Culture; ANIMAL CELLS; ANIMALS; Biological and medical sciences; BIOLOGICAL RECOVERY; BIOLOGICAL REPAIR; CELL CULTURES; Cell lines; Cells; Cells, Cultured; CLONING; Cloning, Molecular; Complementation; Cosmids; Cricetinae; DISEASES; DNA; DNA Damage; DNA HYBRIDIZATION; DNA probes; DNA REPAIR; DNA-CLONING; ELECTROMAGNETIC RADIATION; EXCISION REPAIR; Fundamental and applied biological sciences. Psychology; GENES; Genetic Complementation Test; HAMSTERS; HeLa cells; Humans; Hybrid Cells - radiation effects; HYBRIDIZATION; Kinetics; MAMMALS; MAN; Molecular and cellular biology; Molecular genetics; Mutagenesis. Repair; Nucleic Acid Hybridization; NUCLEIC ACIDS; ORGANIC COMPOUNDS; PRIMATES; RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT; RADIATIONS; RADIOSENSITIVITY; RECOVERY; REPAIR; Restriction Mapping; RODENTS; SKIN DISEASES; Transfection; ULTRAVIOLET RADIATION; Ultraviolet Rays; VERTEBRATES; XERODERMA PIGMENTOSUM; Xeroderma Pigmentosum - genetics; XP CELLS; Cell culture; Molecular hybridization; Excision; Radiolabelling; Nucleotide sequence; Rodentia; Cosmid; Homology; Resistance; Vertebrata; Mammalia; Transfection; Gene; Ultraviolet irradiation; Isolation; Molecular cloning; Repair; Hamster
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.86.18.6997

A hamster DNA repair gene has been isolated by cosmid rescue after two rounds of transfection of an immortalized xeroderma pigmentosum (XP) complementation group D cell line with neomycin-resistance gene (neo)-tagged normal hamster DNA and selection with G418 and ultraviolet irradiation. The functional length of the sequence has been defined as 11.5 kilobase pairs by measurement of the region of overlap between two hamster DNA-containing cosmids, cloned by selection for the integrated neo gene, that are able to confer an increase in resistance to ultraviolet irradiation on two XP-D cell lines but not on an XP-A line. Detailed molecular characterization of the hamster repair gene has revealed no obvious similarities to two human excision repair genes (ERCC1 and ERCC2) that correct repair-defective hamster cells but have no effect on XP cells. Hybridization analyses of normal human and XP cell genomic DNAs and mRNAs, using a cosmid-clone probe from which repeated sequences have been removed, show that homologues are present and expressed in all cases.