Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells.


Aphidicolin specifically inhibits eukaryotic DNA polymerase alpha, ...
Aphidicolin specifically inhibits eukaryotic DNA polymerase alpha, while 2',3'-dideoxythymidine 5'-triphosphate (d2TTP) inhibits DNA polymerase beta and gamma but not alpha. 1-beta-D-Arabinofuranosylcytosine 5'-triphosphate (araCTP) inhibits both DNA polymerase alpha and beta although to a different extent. Here we measured the effects of these inhibitors on repair DNA synthesis of U.V.-irradiated HeLa cells by two different methods. Firstly, aphidicolin, 1-beta-D-arabinofuranosylcytosine (araC, a precursor of araCTP) and 2',3'-dideoxythimidine (d2Thd, a precursor of d2TTP) were added directly to the culture medium. In this case, aphidicolin and araC strongly inhibited replicative DNA synthesis of HeLa cells, and they also inhibited repair synthesis after U.V.-irradiation but to a much lesser extent. In contrast, high concentrations of d2Thd inhibited repair DNA synthesis to a higher extent than replicative DNA synthesis. Secondly, the active form of inhibitor, d2TTP, was microinjection directly into cytoplasm or nuclei or U.V.-irradiated HeLa cells. Microinjection of d2TTP effectively inhibited repair synthesis. The microinjection of d2TTP, into either cytoplasm or nucleus, strongly inhibited replicative synthesis. These results might indicate that multiple DNA polymerases are involved in repair synthesis as well as in replicative synthesis.




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