Molecular cloning of the human nucleotide-excision-repair gene ERCC4

Journal Article · Tue Jul 19 04:00:00 EDT 1994 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.91.15.6855· OSTI ID:86533

Thompson, L H; Brookman, K W; Weber, C A; Salazar, E P ^[1]; Reardon, J T; Sancar, A ^[2]; Deng, Z; Siciliano, M J ^[3]

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1. Lawrence Livermore National Lab., CA (United States)
2. Univ. of North Carolina, Chapel Hill, NC (United States)
3. Univ. of Texas Cancer Center, Houston, TX (United States)

ERCC4 was previously identified in somatic cell hybrids as a human gene that corrects the nucleotide-excision-repair deficiency in mutant hamster cells. The cloning strategy for ERCC4 involved transfection of the repair-deficient hamster cell line UV41 with a human sCos-1 cosmid library derived from chromosome 16. Enhanced UV resistance was seen with one cosmid-library transformant and two secondary transformants of UV41. Cosmid clones carrying a functional ERCC4 gene were isolated from a library of a second transformant by selecting in Escherichia coli for expression of a linked neomycin-resistance gene that was present in the sCos-1 vector. The cosmids mapped to 16p13.13-p13.2, the location assigned to ERCC4 by using somatic cell hybrids. Upon transfection into UV41, six cosmid clones gave partial correction ranging from 30% to 64%, although all appeared to contain the complete gene. The capacity for in vitro excision of thymine dimers from a plasmid by transformant cell extracts correlated qualitatively with enhanced UV resistance.

• Sponsoring Organization: USDOE
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 86533
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 15 Vol. 91; ISSN PNASA6; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
56 BIOLOGY AND MEDICINE
APPLIED STUDIES
DNA
DNA-CLONING
EXCISION REPAIR
GENES
GENETIC RADIATION EFFECTS
HUMAN CHROMOSOME 16
RADIOSENSITIVITY
ULTRAVIOLET RADIATION