Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of saccharomyces cerevisiae and schizosaccharomyces pombe

Journal Article · Fri Oct 01 00:00:00 EDT 1993 · Molecular and Cellular Biology; (United States)

DOI:https://doi.org/10.1128/MCB.13.10.6393· OSTI ID:5852645

MacInnes, M A; Dickson, J A; Hernandez, R R; Lin, G Y; Park, M S; Schauer, S; Reynolds, R J; Strniste, G F ^[1]; Learmonth, D ^[2]; Mudgett, J S ^[3]; Yu, J Y ^[4]

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1. Los Alamos National Lab., NM (United States)
2. West Nistaben, Orkney, Scotland (United Kingdom)
3. Merck Research Laboratories, Rahway, NJ (United States)
4. Univ. of Texas, Austin, TX (United States)

Several human genes related to DNA excision repair (ER) have been isolated via ER cross-species complementation (ERCC) of UV-sensitive CHO cells. The authors have now isolated and characterized cDNAs for the human ERCC5 gene that complement CHO UV135 cells. The ERCC5 mRNA size is about 4.6 kb. Their available cDNA clones are partial length, and no single clone was active for UV135 complementation. When cDNAs were mixed pairwise with a cosmid clone containing an overlapping 5[prime]-end segment of the ERCC5 gene, DNA transfer produced UV-resistant colonies with 60 to 95% correction of UV resistance relative to either a genomic ERCC5 DNA transformant or the CHO AA8 progenitor cells. cDNA-cosmid transformants regained intermediate levels (20 to 45%) of ER-dependent reactivation of a UV-damaged pSVCATgpt reporter plasmid. Their evidence strongly implicates an in situ recombination mechanism in cDNA-cosmid complementation for ER. The complete deduced amino acid sequence of ERCC5 was reconstructed for several cDNA clones encoding a predicted protein of 1,186 amino acids. The ERCC5 protein has extensive sequence similarities, in bipartite domains A and B, to products of RAD repair genes of two yeast, Saccharomyces cerevisiae RAD2 and Schizosaccharomyces pombe rad13. Sequence, structural, and functional data taken together indicate that ERCC5 and its relatives are probable functional homologs. A second locus represented by S. cerevisiae YKL510 and S. pombe rad2 genes is structurally distinct from the ERCC5 locus but retains vestigial A and B domain similarities. Their analyses suggest that ERCC5 is a nuclear-localized protein with one or more highly conserved helix-loop-helix segments within domains A and B. 69 refs., 6 figs., 1 tab.

• OSTI ID: 5852645
• Journal Information: Molecular and Cellular Biology; (United States), Vol. 13:10; ISSN 0270-7306
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CHO CELLS
PHOTOSENSITIVITY
DNA
EXCISION REPAIR
DNA HELICASES
AMINO ACID SEQUENCE
SACCHAROMYCES CEREVISIAE
DNA-CLONING
AMINO ACIDS
DNA REPAIR
DNA SEQUENCING
GENES
GENETIC MAPPING
PROTEINS
ANIMAL CELLS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CARBOXYLIC ACIDS
CLONING
DNA HYBRIDIZATION
ENZYMES
EUMYCOTA
FUNGI
HYBRIDIZATION
MAPPING
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
REPAIR
SACCHAROMYCES
SENSITIVITY
STRUCTURAL CHEMICAL ANALYSIS
YEASTS
550400* - Genetics