DNA single stranded gaps formed during DNA repair synthesis induced by methyl methanesulfonate are filled by sequential action of aphidicolin- and dideoxythymidine sensitive DNA polymerases in HeLa cells |
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Authors: | Il-Seon Park Jong Kun Park Hye Yeong Koh Sang Dai Park |
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Institution: | (1) Department of Zoology, College of Natural Sciences, Seoul National University, 151-742 Seoul, Republic of Korea;(2) Department of Molecular Biology, College of Natural Sciences, Wonkwang University, 570-749 Iri, Republic of Korea;(3) Department of Zoology, College of Natural Sciences, Seoul National University, 151-742 Seoul, Republic of Korea |
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Abstract: | DNA repair synthesis induced by methyl methanesulfonate in preconditioned HeLa cells in which DNA replicative synthesis had been highly suppressed was inhibited by aphidicolin (an inhibitor of DNA polymerases and ) and dideoxythymidine (ddThR, an inhibitor of DNA polymerase ). Incomplete repair patches sensitive to exonuclease III were accumulated in the presence of aphidicolin while not in the presence of ddThR. These patches were comopleted by the combined action of Klenow fragment and T4 DNA ligase, indicating that the single-stranded gaps were formed during the repair synthesis. Moreover, ddThR had little effect on the repair synthesis in the presence of aphidicolin. Thus, the results suggest that the single-stranded gaps may be sealed first by aphidicolin-sensitive polymerase followed by ddThR-sensitive DNA polymerase on the same site of the repair patch.Abbreviations ddThR
dideoxythymidine
- MMS
methyl methanesulfonate
- dNTP
deoxynucleoside triphosphate |
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Keywords: | aphidicolin ddThR DNA strand breaks excision repair MMS |
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