Targeting single-strand DNA breaks for DNA repairстатьяТезисы
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Дата последнего поиска статьи во внешних источниках: 16 января 2019 г.
Аннотация:Single-strand DNA breaks (SSBs) is the prevalent form ofendogenous DNA damage in the living cells. They are frequentlycaused by reactive oxygen species, which directly disintegratebackbone sugars and often produce damaged 3’ and 5’ terminisurrounding the break. SSB repair (SSBR) often engages proteinsdedicated to base excision repair (BER) and is considered as itssub-pathway. In higher eukaryotes one of the major proteinsdetecting such breaks and orchestrating SSBR is poly(ADP-ribose) polymerase 1 (PARP-1). Elongating RNA polymerase II(RNAP II) also participates in detection of these lesions. SSBs intemplate strand (TS-SSBs) of a transcribed DNA cause arrest ofsome RNAPs with efficiencies varying significantly depending onthe 5’ and 3’ backbone end chemistries at the break and the pres-ence or absence of adjacent abasic sites. In contrast, SSBs innon-transcribed strand (NT-SSBs) are shown to inhibit RNAPsonly during transcription through a nucleosome. The effect ofTS-SSBs on transcript elongation byE. coliRNAP and RNAPII was not yet investigated in details. In the present work weestablished anin vitroexperimental system for investigation oftranscription of histone-free and nucleosomal DNA in presenceof randomly or uniquely introduced TS-SSBs. The effect of TS-SSBs on elongation byE. coliRNAP was studied using the sys-tem. Damaged templates were transcribedin vitrobyE. coliRNAP, and efficiency of elongation through the lesion andenzyme stalling were evaluated. The system can be adapted fortranscription by RNAP II and will be used to study the role ofDNA repair machinery during ongoing transcription and toreveal how PARP-1 affects elongation on the damaged templates.This work was supported by RFBR grant No. 17-54-33045 andThe Russian Federation Presidential Grant for Young ScientistsNo MK-2377.2017.4 (Agreement of the Ministry of Education ofthe Russian Federation No 14.W01.17.2377-MK).