Sequential binding of DNA repair proteins RPA and ERCC1 to XPA in vitro.
AUTOR(ES)
Saijo, M
RESUMO
Recent studies have shown that many proteins are involved in the early steps of nucleotide excision repair and that there are some interactions between nucleotide excision repair proteins, suggesting that these interactions are important in the reaction mechanism. The xeroderma pigmentosum group A protein (XPA) was shown to bind to the replication protein A (RPA) or the excision repair cross complementing rodent repair deficiency group 1 protein (ERCC1), and these interactions might be involved in the damage-recognition and/or incision steps, of nucleotide excision repair. Here we show that the XPA regions required for the binding to the 70 and 34 kDa subunits of RPA are located in the middle and on N-terminal regions of XPA, respectively. These regions do not overlap with the ERCC1-binding region of XPA, and a ternary protein complex of RPA, XPA and ERCC1 was detected in vitro. In addition, using the surface plasmon resonance biosensor, the binding of RPA and ERCC1 to XPA was investigated. The dissociation constants (KD) of RPA and ERCC1 with XPA were 1.9 x 10(-8 )and 2.5 x 10(-7) M, respectively. Moreover, our results suggest the sequential binding of RPA and ERCC1 to XPA.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=146300Documentos Relacionados
- Specific association between the human DNA repair proteins XPA and ERCC1.
- Mutations in XPA that prevent association with ERCC1 are defective in nucleotide excision repair.
- Mapping of interaction domains between human repair proteins ERCC1 and XPF.
- Activity of individual ERCC1 and XPF subunits in DNA nucleotide excision repair
- Formation of a ternary complex by human XPA, ERCC1, and ERCC4(XPF) excision repair proteins.