Localization of xeroderma pigmentosum group A protein and replication protein A on damaged DNA in nucleotide excision repair

The interaction of xeroderma pigmentosum group A protein (XPA) and replication protein A (RPA) with damaged DNA in nucleotide excision repair (NER) was studied using model dsDNA and bubble-DNA structure with 5-{3-[6-(carboxyamido-fluoresceinyl)amidocapromoyl]allyl}-dUMP lesions in one strand and con...

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Bibliographic Details
Published in:Nucleic acids research 2010-12, Vol.38 (22), p.8083-8094
Main Authors: Krasikova, Yuliya S, Rechkunova, Nadejda I, Maltseva, Ekaterina A, Petruseva, Irina O, Lavrik, Olga I
Format: Article
Language:English
Subjects:
Deoxyribonucleases - metabolism
DNA - chemistry
DNA - metabolism
DNA Damage
DNA Footprinting
DNA Repair
Genome Integrity, Repair and
Replication Protein A - analysis
Replication Protein A - metabolism
Xeroderma Pigmentosum Group A Protein - analysis
Xeroderma Pigmentosum Group A Protein - metabolism
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Summary:The interaction of xeroderma pigmentosum group A protein (XPA) and replication protein A (RPA) with damaged DNA in nucleotide excision repair (NER) was studied using model dsDNA and bubble-DNA structure with 5-{3-[6-(carboxyamido-fluoresceinyl)amidocapromoyl]allyl}-dUMP lesions in one strand and containing photoreactive 5-iodo-dUMP residues in defined positions. Interactions of XPA and RPA with damaged and undamaged DNA strands were investigated by DNA-protein photocrosslinking and gel shift analysis. XPA showed two maximums of crosslinking intensities located on the 5'-side from a lesion. RPA mainly localized on undamaged strand of damaged DNA duplex and damaged bubble-DNA structure. These results presented for the first time the direct evidence for the localization of XPA in the 5'-side of the lesion and suggested the key role of XPA orientation in conjunction with RPA binding to undamaged strand for the positioning of the NER preincision complex. The findings supported the mechanism of loading of the heterodimer consisting of excision repair cross-complementing group 1 and xeroderma pigmentosum group F proteins by XPA on the 5'-side from the lesion before damaged strand incision. Importantly, the proper orientation of XPA and RPA in the stage of preincision was achieved in the absence of TFIIH and XPG.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/gkq649