Abstract: Dehydroaltenusin is a selective inhibitor of mammalian DNA polymerase α (pol α) from a fungus (Alternaria tennuis). We have designed, synthesized, and characterized a derivative of dehydroaltenusin conjugated with a C12-alkyl side chain (dehydroaltenusin-C12 [C12]). C12 was the strongest pol α inhibitor in vitro. We introduced C12 into NIH3T3 cells with the help of a hypotonic shift, that is, a transient exposure of cultured cells in hypotonic buffer with small molecules which can not penetrate cells. The cells that took in C12 by hypotonic shift showed cell growth inhibition. At a low concentration (5 μM), DNA replication was inhibited and several large replication protein A (RPA) foci, which is different from dUTP foci. Furthermore, when C12 was incubated with aphidicolin, RPA foci were not observed in cells. Finally, these findings suggest that C12 inhibited DNA replication through pol α inhibition, and generated single-stranded DNA, resulted in uncoupling of the leading strand and lagging strand synthesis. These findings suggest that C12 could be more interesting as a molecule probe or anticancer agent than aphidicolin. C12 might provide novel markers for the development of antiproliferative drugs.
Keywords: Dehydroaltenusin-C12 (C12); DNA polymerase (DNA-directed DNA polymerase [E.C. 22.214.171.124]; pol) α; Enzyme inhibitor; DNA replication; Replication fork uncoupling; Hypotonic shift; Molecule probe; Anticancer drug
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Kuriyama, I.; Mizuno, T.; Fukudome, K.; Kuramochi, K.; Tsubaki, K.; Usui, T.; Imamoto, N.; Sakaguchi, K.; Sugawara, F.; Yoshida, H.; Mizushina, Y. Effect of Dehydroaltenusin-C12 Derivative, a Selective DNA Polymerase α Inhibitor, on DNA Replication in Cultured Cells. Molecules 2008, 13, 2948-2961.
Kuriyama I, Mizuno T, Fukudome K, Kuramochi K, Tsubaki K, Usui T, Imamoto N, Sakaguchi K, Sugawara F, Yoshida H, Mizushina Y. Effect of Dehydroaltenusin-C12 Derivative, a Selective DNA Polymerase α Inhibitor, on DNA Replication in Cultured Cells. Molecules. 2008; 13(12):2948-2961.
Kuriyama, Isoko; Mizuno, Takeshi; Fukudome, Keishi; Kuramochi, Kouji; Tsubaki, Kazunori; Usui, Takeo; Imamoto, Naoko; Sakaguchi, Kengo; Sugawara, Fumio; Yoshida, Hiromi; Mizushina, Yoshiyuki. 2008. "Effect of Dehydroaltenusin-C12 Derivative, a Selective DNA Polymerase α Inhibitor, on DNA Replication in Cultured Cells." Molecules 13, no. 12: 2948-2961.