Abstract: first_pagesettingsOrder Article Reprints Open AccessArticle Correlated Target Search by Vaccinia Virus Uracil–DNA Glycosylase, a DNA Repair Enzyme and a Processivity Factor of Viral Replication Machinery by Evgeniia A. Diatlova 1,Grigory V. Mechetin 1,Anna V. Yudkina 1,Vasily D. Zharkov 2,Natalia A. Torgasheva 1,Anton V. Endutkin 1ORCID,Olga V. Shulenina 3,Andrey L. Konevega 3ORCID,Irina P. Gileva 4,Sergei N. Shchelkunov 4ORCID andDmitry O. Zharkov 1,5,*ORCID 1 SB RAS Institute of Chemical Biology and Fundamental Medicine, 8 Lavrentieva Ave., 630090 Novosibirsk, Russia 2 Biology Department, Tomsk State University, 634050 Tomsk, Russia 3 NRC “Kurchatov Institute”—B. P. Konstantinov Petersburg Nuclear Physics Institute, Leningrad Region, 188300 Gatchina, Russia 4 State Research Center of Virology and Biotechnology Vector, Novosibirsk Region, 630559 Koltsovo, Russia 5 Department of Natural Sciences, Novosibirsk State University, 2 Pirogova St., 630090 Novosibirsk, Russia * Author to whom correspondence should be addressed. Int. J. Mol. Sci. 2023, 24(11), 9113; https://doi.org/10.3390/ijms24119113 Received: 20 April 2023 / Revised: 13 May 2023 / Accepted: 21 May 2023 / Published: 23 May 2023 (This article belongs to the Special Issue Recent Advances in Genome Maintenance Studies) Download Browse Figures Versions Notes Abstract The protein encoded by the vaccinia virus D4R gene has base excision repair uracil–DNA N-glycosylase (vvUNG) activity and also acts as a processivity factor in the viral replication complex. The use of a protein unlike PolN/PCNA sliding clamps is a unique feature of orthopoxviral replication, providing an attractive target for drug design. However, the intrinsic processivity of vvUNG has never been estimated, leaving open the question whether it is sufficient to impart processivity to the viral polymerase. Here, we use the correlated cleavage assay to characterize the translocation of vvUNG along DNA between two uracil residues. The salt dependence of the correlated cleavage, together with the similar affinity of vvUNG for damaged and undamaged DNA, support the one-dimensional diffusion mechanism of lesion search. Unlike short gaps, covalent adducts partly block vvUNG translocation. Kinetic experiments show that once a lesion is found it is excised with a probability ~0.76. Varying the distance between two uracils, we use a random walk model to estimate the mean number of steps per association with DNA at ~4200, which is consistent with vvUNG playing a role as a processivity factor. Finally, we show that inhibitors carrying a tetrahydro-2,4,6-trioxopyrimidinylidene moiety can suppress the processivity of vvUNG.

Cite: Dyatlova E. , Mechetin G.V. , Yudkina A.V. , Zharkov D.O. , Torgasheva N.A. , Endutkin A.V. , Shulenina O.V. , Konevega A.L. , Gileva I.P. , Shchelkunov S.N. , Zharkov D.O.
Correlated target search by vaccinia virus uracil–DNA glycosylase, a DNA repair enzyme and a processivity factor of viral replication machinery
International Journal of Molecular Sciences. 2023. V.24. N11. 9113 . DOI: 10.3390/ijms24119113 WOS Scopus РИНЦ OpenAlex

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Web of science:	WOS:001005116400001
Scopus:	2-s2.0-85161586267
Elibrary:	7298065
OpenAlex:	W4378078112

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