Mapping of interaction domains between human repair proteins ERCC1 and XPF
ERCC1-XPF is a heterodimeric protein complexinvolved in nucleotide excision repair and recombinational processes. Like its homologous complex in Saccharomyces cerevisiae , Rad10-Rad1, it acts as a structure-specific DNA endonuclease, cleaving at duplex-single-stranded DNA junctions. In repair, ERCC1-XPF and Rad10-Rad1 make an incision on the the 5'-side of the lesion. No humans with a defect in the ERCC1 subunit of this protein complex have been identified and ERCC1-deficient mice suffer from severe developmental problems and signs of premature aging on top of a repair-deficient phenotype. Xeroderma pigmentosum group F patients carry mutations in the XPF subunit and generally show the clinical symptoms of mild DNA repair deficiency. All XP-F patients examined demonstrate reduced levels of XPF and ERCC1 protein, suggesting that proper complex formation is required for stability of the two proteins. To better understand the molecular and clinical consequences of mutations in the ERCC1-XPF complex, we decided to map the interaction domains between the two subunits. The XPF-binding domain comprises C-terminal residues 224-297 of ERCC1. Intriguingly, this domain resides outside the region of homology with its yeast Rad10 counterpart. The ERCC1-binding domain in XPF maps to C-terminal residues 814-905. ERCC1-XPF complex formation is established by a direct interaction between these two binding domains. A mutation from an XP-F patient that alters the ERCC1-binding domain in XPF indeed affects complex formation with ERCC1.
|*DNA Repair, *Saccharomyces cerevisiae Proteins, Animals, Comparative Study, DNA-Binding Proteins/*chemistry/genetics/*metabolism, Endonucleases/metabolism, Fungal Proteins/metabolism, Humans, Macromolecular Substances, Mammals, Mice, Mice, Knockout, Polymerase Chain Reaction, Protein Biosynthesis, Proteins/*chemistry/genetics/*metabolism, Research Support, Non-U.S. Gov't, Sequence Deletion, Transcription, Genetic|
|Nucleic Acids Research|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
de Laat, W.L, Sijbers, A.M, Odijk, H, Hoeijmakers, J.H.J, & Jaspers, N.G.J. (1998). Mapping of interaction domains between human repair proteins ERCC1 and XPF. Nucleic Acids Research. Retrieved from http://hdl.handle.net/1765/8893