Two interaction surfaces between XPA and RPA organize the preincision complex in nucleotide excision repair

Kim, Mihyun; Kim, Hyun-Suk; D’Souza, Areetha; Gallagher, Kaitlyn; Jeong, Eunwoo; Topolska-Woś, Agnieszka; Ogorodnik Le Meur, Kateryna; Tsai, Chi-Lin; Tsai, Miaw-Sheue; Kee, Minyong; Tainer, John A.; Yeo, Jung-Eun; Chazin, Walter J.; Schärer, Orlando D.

doi:10.1073/pnas.2207408119

Title: Two interaction surfaces between XPA and RPA organize the preincision complex in nucleotide excision repair

Journal Article · 15 August 2022 · Proceedings of the National Academy of Sciences of the United States of America

DOI: https://doi.org/10.1073/pnas.2207408119 · OSTI ID:1969342

^[1]; Kim, Hyun-Suk ^[2]; D’Souza, Areetha ^[3]; Gallagher, Kaitlyn ^[3];

^[2]; Topolska-Woś, Agnieszka ^[3]; Ogorodnik Le Meur, Kateryna ^[3];

^[4]; Tsai, Miaw-Sheue ^[5]; Kee, Minyong ^[2];

^[4]; Yeo, Jung-Eun ^[2];

^[3];

^[6]

Inst. for Basic Science (Korea, Republic of); Ulsan National Institute of Science and Technology (UNIST), Ulsan (Korea, Republic of)
Inst. for Basic Science (Korea, Republic of)
Vanderbilt Univ., Nashville, TN (United States)
Univ. of Texas, Houston, TX (United States)
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Inst. for Basic Science (Korea, Republic of); Ulsan National Institute of Science and Technology (UNIST), Ulsan (Korea, Republic of); Vanderbilt Univ., Nashville, TN (United States)

The xeroderma pigmentosum protein A (XPA) and replication protein A (RPA) proteins fulfill essential roles in the assembly of the preincision complex in the nucleotide excision repair (NER) pathway. We have previously characterized the two interaction sites, one between the XPA N-terminal (XPA-N) disordered domain and the RPA32 C-terminal domain (RPA32C), and the other with the XPA DNA binding domain (DBD) and the RPA70AB DBDs. Here, we show that XPA mutations that inhibit the physical interaction in either site reduce NER activity in biochemical and cellular systems. Combining mutations in the two sites leads to an additive inhibition of NER, implying that they fulfill distinct roles. Our data suggest a model in which the interaction between XPA-N and RPA32C is important for the initial association of XPA with NER complexes, while the interaction between XPA DBD and RPA70AB is needed for structural organization of the complex to license the dual incision reaction. Integrative structural models of complexes of XPA and RPA bound to single-stranded/double-stranded DNA (ss/dsDNA) junction substrates that mimic the NER bubble reveal key features of the architecture of XPA and RPA in the preincision complex. Most critical among these is that the shape of the NER bubble is far from colinear as depicted in current models, but rather the two strands of unwound DNA must assume a U-shape with the two ss/dsDNA junctions localized in close proximity. Our data suggest that the interaction between XPA and RPA70 is key for the organization of the NER preincision complex.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: Korean Institute for Basic Science; National Cancer Institute (NCI); National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1969342

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 34 Vol. 119; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
DNA repair
nucleotide excision repair
replication protein A
xeroderma pigmentosum protein A

Title: Two interaction surfaces between XPA and RPA organize the preincision complex in nucleotide excision repair

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