Physical and functional interaction between the XPF/ERCC1 endonuclease and hRad52.
The XPF/ERCC1 heterodimer is a DNA structure-specific endonuclease that participates in nucleotide excision repair and homology-dependent recombination reactions, including DNA single strand annealing and gene targeting. Here we show that XPF/ERCC1 is stably associated with hRad52, a recombinational repair protein, in human cell-free extracts and that these factors interact directly via the N-terminal domain of hRad52 and the XPF protein. Complex formation between hRad52 and XPF/ERCC1 concomitantly stimulates the DNA structure-specific endonuclease activity of XPF/ERCC1 and attenuates the DNA strand annealing activity of hRad52. Our results reveal a novel role for hRad52 as a subunit of a DNA structure-specific endonuclease and are congruent with evidence implicating both hRad52 and XPF/ERCC1 in a number of homologous recombination reactions. We propose that the ternary complex of hRad52 and XPF/ERCC1 is the active species that processes recombination intermediates generated during the repair of DNA double strand breaks and in homology-dependent gene targeting events.
Motycka
Teresa A
TA
Molecular Medicine Graduate Program, Institute of Biotechnology, The University of Texas Health Science Center, San Antonio, Texas 78245, USA.
Bessho
Tadayoshi
T
Post
Sean M
SM
Sung
Patrick
P
Tomkinson
Alan E
AE
D004247:DNA / Q000378:metabolism; D004260:DNA Repair / Q000502:physiology; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004720:Endonucleases; D004789:Enzyme Activation; D006367:HeLa Cells; D006801:Humans; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D051721:Rad52 DNA Repair and Recombination Protein; D011995:Recombination, Genetic / Q000502:physiology
D004268:DNA-Binding Proteins; D011506:Proteins; C497194:RAD52 protein, human; D051721:Rad52 DNA Repair and Recombination Protein; C095888:xeroderma pigmentosum group F protein; D004247:DNA; C071447:ERCC1 protein, human; D004720:Endonucleases