Petasiphenol: a DNA polymerase lambda inhibitor.
Petasiphenol, a bio-antimutagen isolated from a Japanese vegetable, Petasites japonicus, selectively inhibits the activities of mammalian DNA polymerase lambda (pol lambda) in vitro. The compound did not influence the activities of replicative DNA polymerases such as alpha, delta, and epsilon but also showed no effect even on the pol beta activity, the three-dimensional structure of which is thought to be highly similar to pol lambda. The inhibitory effect of petasiphenol on intact pol lambda including the BRCA1 C-terminus (BRCT) domain was dose-dependent, and 50% inhibition was observed at a concentration of 7.8 microM. The petasiphenol-induced inhibition of the pol lambda activity was noncompetitive with respect to both the DNA template-primer and the dNTP substrate. Petasiphenol did not only inhibit the activity of the truncated pol lambda including the pol beta-like core, in which the BRCT motif was deleted in its N-terminal region. BIAcore analysis demonstrated that petasiphenol bound selectively to the N-terminal domain of pol lambda but did not bind to the C-terminal region. On the basis of these results, the pol lambda inhibitory mechanism of petasiphenol is discussed.
Mizushina
Yoshiyuki
Y
Laboratory of Food and Nutritional Sciences, Department of Nutritional Science, and High Technology Research Center, Kobe-Gakuin University, Nishi-ku, Kobe, Hyogo 651-2180, Japan. mizushin@nutr.kobegakuin.ac.jp
Kamisuki
Shinji
S
Kasai
Nobuyuki
N
Ishidoh
Tomomi
T
Shimazaki
Noriko
N
Takemura
Masaharu
M
Asahara
Hitomi
H
Linn
Stuart
S
Yoshida
Shonen
S
Koiwai
Osamu
O
Sugawara
Fumio
F
Yoshida
Hiromi
H
Sakaguchi
Kengo
K
D000595:Amino Acid Sequence; D000818:Animals; D016587:Antimutagenic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D019313:BRCA1 Protein / Q000737:chemistry; D001665:Binding Sites; D002109:Caffeic Acids / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D002417:Cattle; D003198:Computer Simulation; D019951:DNA Polymerase beta / Q000037:antagonists & inhibitors* / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D036901:Petasites / Q000737:chemistry; D017434:Protein Structure, Tertiary; D051381:Rats; D017386:Sequence Homology, Amino Acid
D016587:Antimutagenic Agents; D019313:BRCA1 Protein; D002109:Caffeic Acids; D004791:Enzyme Inhibitors; D010446:Peptide Fragments; C079021:petasiphenol; C415367:DNA polymerase beta2; D019951:DNA Polymerase beta