Inhibition of topoisomerase I by NAD and enhancement of cytotoxicity of MMS by inhibitors of poly(ADP-ribose) polymerase in Saccharomyces cerevisiae.
The activity of DNA topoisomerase I present in the nuclear extract of yeast, Saccharomyces cerevisiae, was inhibited by additions of NAD, the substrate of poly (ADP-ribose) polymerase. This NAD-inhibited topoisomerase activity was restored to the normal level in a dose-dependent manner by adding 3-aminobenzamide (3-AB), an inhibitor of the polymerase. The 3-AB sensitive polymerase enzyme activity, as determined by the rate of incorporation of the radiolabelled NAD in permeabilized cells, increased by treatment of cells with methyl methanesulfonate (MMS) in a dose-dependent manner. While the additions of MMS increased the polymerase activity, it has caused a decrease in cell survival. However, this cell killing activity of MMS was markedly potentiated by adding benzamide, another inhibitor of polymerase. Thus, these results suggest that the mode of modification of nuclear proteins by altering the poly(ADP-ribosylation) in S. cerevisiae resembles with those observed in mammalian cells.
Park
J K
JK
Department of Molecular Biology, Wonkwang University, Seoul, Republic of Korea.
Kim
W J
WJ
Park
Y S
YS
Choi
H S
HS
Yu
J E
JE
Han
D M
DM
Park
S D
SD
D001549:Benzamides / Q000494:pharmacology; D008741:Methyl Methanesulfonate / Q000494:pharmacology*; D009243:NAD / Q000378:metabolism*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D059004:Topoisomerase I Inhibitors
D001549:Benzamides; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D059004:Topoisomerase I Inhibitors; D009243:NAD; C025160:3-aminobenzamide; D008741:Methyl Methanesulfonate; D011065:Poly(ADP-ribose) Polymerases