TY - JOUR

T1 - Does the XPA–FEN1 Interaction Concern to Nucleotide Excision Repair or Beyond?

AU - Krasikova, Yuliya S.

AU - Maltseva, Ekaterina A.

AU - Khodyreva, Svetlana N.

AU - Evdokimov, Alexey N.

AU - Rechkunova, Nadejda I.

AU - Lavrik, Olga I.

N1 - This work was supported by the Russian Science Foundation (grant numbers 21-64-00017 and 19-74-10056, for fluorescently labeled XPA preparation and analysis) and by the Ministry of Science and Higher Education of Russian Federation (project number 121031300041-4, for DNA duplex preparation and protein purification).

PY - 2024/7

Y1 - 2024/7

N2 - Nucleotide excision repair (NER) is the most universal repair pathway, which removes a wide range of DNA helix-distorting lesions caused by chemical or physical agents. The final steps of this repair process are gap-filling repair synthesis and subsequent ligation. XPA is the central NER scaffolding protein factor and can be involved in post-incision NER stages. Replication machinery is loaded after the first incision of the damaged strand that is performed by the XPF–ERCC1 nuclease forming a damaged 5′-flap processed by the XPG endonuclease. Flap endonuclease I (FEN1) is a critical component of replication machinery and is absolutely indispensable for the maturation of newly synthesized strands. FEN1 also contributes to the long-patch pathway of base excision repair. Here, we use a set of DNA substrates containing a fluorescently labeled 5′-flap and different size gap to analyze possible repair factor–replication factor interactions. Ternary XPA–FEN1–DNA complexes with each tested DNA are detected. Furthermore, we demonstrate XPA–FEN1 complex formation in the absence of DNA due to protein–protein interaction. Functional assays reveal that XPA moderately inhibits FEN1 catalytic activity. Using fluorescently labeled XPA, formation of ternary RPA–XPA–FEN1 complex, where XPA accommodates FEN1 and RPA contacts simultaneously, can be proposed. We discuss possible functional roles of the XPA–FEN1 interaction in NER related DNA resynthesis and/or other DNA metabolic processes where XPA can be involved in the complex with FEN1.

AB - Nucleotide excision repair (NER) is the most universal repair pathway, which removes a wide range of DNA helix-distorting lesions caused by chemical or physical agents. The final steps of this repair process are gap-filling repair synthesis and subsequent ligation. XPA is the central NER scaffolding protein factor and can be involved in post-incision NER stages. Replication machinery is loaded after the first incision of the damaged strand that is performed by the XPF–ERCC1 nuclease forming a damaged 5′-flap processed by the XPG endonuclease. Flap endonuclease I (FEN1) is a critical component of replication machinery and is absolutely indispensable for the maturation of newly synthesized strands. FEN1 also contributes to the long-patch pathway of base excision repair. Here, we use a set of DNA substrates containing a fluorescently labeled 5′-flap and different size gap to analyze possible repair factor–replication factor interactions. Ternary XPA–FEN1–DNA complexes with each tested DNA are detected. Furthermore, we demonstrate XPA–FEN1 complex formation in the absence of DNA due to protein–protein interaction. Functional assays reveal that XPA moderately inhibits FEN1 catalytic activity. Using fluorescently labeled XPA, formation of ternary RPA–XPA–FEN1 complex, where XPA accommodates FEN1 and RPA contacts simultaneously, can be proposed. We discuss possible functional roles of the XPA–FEN1 interaction in NER related DNA resynthesis and/or other DNA metabolic processes where XPA can be involved in the complex with FEN1.

KW - DNA repair

KW - DNA replication

KW - FEN1

KW - XPA

KW - nucleotide excision repair

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85199606696&origin=inward&txGid=64816eb46286dcb8ac9973cd3d813552

UR - https://www.mendeley.com/catalogue/f62f6248-b1c7-302d-910f-2186cc34bc03/

U2 - 10.3390/biom14070814

DO - 10.3390/biom14070814

M3 - Article

C2 - 39062528

VL - 14

JO - Biomolecules

JF - Biomolecules

SN - 2218-273X

IS - 7

M1 - 814

ER -