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PMID:8197175

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Citation

Park, CH and Sancar, A (1994) Formation of a ternary complex by human XPA, ERCC1, and ERCC4(XPF) excision repair proteins. Proc. Natl. Acad. Sci. U.S.A. 91:5017-21

Abstract

The xeroderma pigmentosum complementation group A (XP-A) protein, XPA, has recently been expressed in Escherichia coli in a soluble and fully functional form. An affinity column was prepared by linking the XPA protein to a solid support. When HeLa cell-free extract capable of excision repair was applied to the column, > 99.9% of the proteins were in the flow-through. However, the flow-through fraction lacked excision activity. The activity was restored by adding the high salt (1 M KCl) eluate of the column to the flow-through fraction. The XPA protein-bound fraction was tested for specific proteins by an in vitro complementation assay with a panel of cell-free extracts from DNA repair-deficient human and rodent cell lines. The XPA-bound fraction complemented cell-free extracts of excision repair cross-complementing 1 (ERCC-1), ERCC-4 (XP-F), and XP-A mutants. We conclude that the XPA damage recognition protein makes a ternary complex with the ERCC1/ERCC4(XPF) heterodimer with a potential nuclease function.

Links

PubMed PMC43921

Keywords

Animals; Cell-Free System; Chromatography, Affinity; Cloning, Molecular; DNA Damage; DNA Repair; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/metabolism; Endonucleases; Escherichia coli; Exonucleases/metabolism; Genetic Complementation Test; HeLa Cells; Humans; Protein Binding; Proteins/chemistry; Proteins/metabolism; Xeroderma Pigmentosum Group A Protein

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:ERCC1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P23025

F

Seeded From UniProt

complete

HUMAN:ERCC1

part_of

GO:0000109: nucleotide-excision repair complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:XPA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P07992

F

Seeded From UniProt

complete

HUMAN:XPA

enables

GO:0042803: protein homodimerization activity

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P23025

F

Seeded From UniProt

complete


See also

References

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