Reconstitution of human excision nuclease with recombinant XPF-ERCC1 complex
- PMID: 9013642
- DOI: 10.1074/jbc.272.6.3833
Reconstitution of human excision nuclease with recombinant XPF-ERCC1 complex
Abstract
The human XPF-ERCC1 protein complex is one of several factors known to be required for general nucleotide excision repair. Genetic data indicate that both proteins of this complex are necessary for the repair of interstrand cross-links, perhaps via recombination. To determine whether XPF-ERCC1 completes a set of six proteins that are sufficient to carry out excision repair, the human XPF and ERCC1 cDNAs were coexpressed in Sf21 insect cells from a baculovirus vector. The purified complex contained the anticipated 5' junction-specific endonuclease activity that is stimulated through a direct interaction between XPF and replication protein A (RPA). The recombinant complex also complemented extracts of XP-F cells and Chinese hamster ovary mutants assigned to complementation groups 1, 4, and 11. Furthermore, reconstitution of the human excision nuclease was observed with a mixture of five repair factors (XPA, XPC, XPG, TFIIH, and RPA) and the recombinant XPF-ERCC1, thus verifying that no additional protein factors are needed for the specific dual incisions characteristic of human excision repair.
Similar articles
-
Replication protein A confers structure-specific endonuclease activities to the XPF-ERCC1 and XPG subunits of human DNA repair excision nuclease.J Biol Chem. 1996 May 10;271(19):11047-50. doi: 10.1074/jbc.271.19.11047. J Biol Chem. 1996. PMID: 8626644
-
Activity of individual ERCC1 and XPF subunits in DNA nucleotide excision repair.Nucleic Acids Res. 2001 Feb 15;29(4):872-9. doi: 10.1093/nar/29.4.872. Nucleic Acids Res. 2001. PMID: 11160918 Free PMC article.
-
DNA structural elements required for ERCC1-XPF endonuclease activity.J Biol Chem. 1998 Apr 3;273(14):7835-42. doi: 10.1074/jbc.273.14.7835. J Biol Chem. 1998. PMID: 9525876
-
The ERCC1 and ERCC4 (XPF) genes and gene products.Gene. 2015 Sep 15;569(2):153-61. doi: 10.1016/j.gene.2015.06.026. Epub 2015 Jun 12. Gene. 2015. PMID: 26074087 Free PMC article. Review.
-
Interstrand crosslink repair: can XPF-ERCC1 be let off the hook?Trends Genet. 2008 Feb;24(2):70-6. doi: 10.1016/j.tig.2007.11.003. Epub 2008 Jan 14. Trends Genet. 2008. PMID: 18192062 Review.
Cited by
-
Removal of reactive oxygen species-induced 3'-blocked ends by XPF-ERCC1.Chem Res Toxicol. 2011 Nov 21;24(11):1876-81. doi: 10.1021/tx200221j. Epub 2011 Oct 18. Chem Res Toxicol. 2011. PMID: 22007867 Free PMC article.
-
Rodent UV-sensitive mutant cell lines in complementation groups 6-10 have normal general excision repair activity.Nucleic Acids Res. 1997 Mar 1;25(5):1015-21. doi: 10.1093/nar/25.5.1015. Nucleic Acids Res. 1997. PMID: 9023113 Free PMC article.
-
The ERCC1/XPF endonuclease is required for completion of homologous recombination at DNA replication forks stalled by inter-strand cross-links.Nucleic Acids Res. 2009 Oct;37(19):6400-13. doi: 10.1093/nar/gkp705. Epub 2009 Aug 27. Nucleic Acids Res. 2009. PMID: 19713438 Free PMC article.
-
The Intriguing Mystery of RPA Phosphorylation in DNA Double-Strand Break Repair.Genes (Basel). 2024 Jan 27;15(2):167. doi: 10.3390/genes15020167. Genes (Basel). 2024. PMID: 38397158 Free PMC article. Review.
-
Initiation of DNA interstrand cross-link repair in mammalian cells.Environ Mol Mutagen. 2010 Jul;51(6):604-24. doi: 10.1002/em.20559. Environ Mol Mutagen. 2010. PMID: 20658650 Free PMC article. Review.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases
Research Materials
