New DNA plasmid model for studying DNA mismatch repair response to the G4 structure

Pavlova, A.V.; Dolinnaya, N.G.; Zvereva, M.I.; Kubareva, E.A.; Monakhova, M.V.

Авторы: Pavlova A.V., Dolinnaya N.G., Zvereva M.I., Kubareva E.A., Monakhova M.V.
Журнал: International Journal of Molecular Sciences
Том: 24
Номер: 2
Год издания: 2023
Издательство: MDPI
Местоположение издательства: Basel, Switzerland
Первая страница: 1061
DOI: 10.3390/ijms24021061
Аннотация: G-quadruplexes (G4s), the most widely studied alternative DNA structures, are implicatedin the regulation of the key cellular processes. In recent years, their involvement in DNA repairmachinery has become the subject of intense research. Here, we evaluated the effect of G4 onthe prokaryotic DNA mismatch repair (MMR) pathway from two bacterial sources with differentmismatch repair mechanisms. The G4 folding, which competes with the maintenance of double-stranded DNA, is known to be controlled by numerous opposing factors. To overcome the kineticbarrier of G4 formation, we stabilized a parallel G4 formed by the d(GGGT)4 sequence in a DNAplasmid lacking a fragment complementary to the G4 motif. Unlike commonly used isolated G4structures, our plasmid with an embedded stable G4 structure contained elements, such as a MutHcleavage site, required to initiate the repair process. G4 formation in the designed construct wasconfirmed by Taq polymerase stop assay and dimethyl sulfate probing. The G4-carrying plasmid,together with control ones (lacking a looped area or containing unstructured d(GT)8insert instead of the G4 motif), were used as new type models to answer the question of whether G4 formation interferes with DNA cleavage as a basic function of MMR.
Добавил в систему: Кубарева Елена Александровна

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New DNA plasmid model for studying DNA mismatch repair response to the G4 structureстатья Исследовательская статья Электронная публикация

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