Flavones inhibit the hexameric replicative helicase RepA.
Helicases couple the hydrolysis of nucleoside triphosphates (NTPs) to the unwinding of double-stranded nucleic acids and are essential in DNA metabolism. Thus far, no inhibitors are known for helicases except heliquinomycin isolated from Streptomyces sp. As the three-dimensional structure of the hexameric replicative DNA helicase RepA encoded by the broad host-range plasmid RSF1010 is known, this protein served as a model helicase to search for inhibitory compounds. The commercially available flavone derivatives luteolin, morin, myricetin and dimyricetin (an oxidation product of myricetin) inhibited the ATPase and double-stranded DNA unwinding activities of RepA. Dimyricetin was the most effective inhibitor for both activities. Single-stranded DNA-dependent RepA ATPase activity is inhibited non-competitively by all four compounds. This finding contrasts the inhibition of phosphoinositide 3-kinase by flavones that fit into the ATP binding pocket of this enzyme. Myricetin also inhibited the growth of a Gram-positive and a Gram-negative bacterial species. As we found other hexameric and non-hexameric prokaryotic helicases to be differentially sensitive to myricetin, flavones may provide substructures for the design of molecules helpful for unraveling the mechanism of helicase action and of novel pharmacologically useful molecules.
Xu
H
H
Institut für Kristallographie, Freie Universität Berlin, Takustrasse 6, D-14195 Berlin, Germany.
Ziegelin
G
G
Schröder
W
W
Frank
J
J
Ayora
S
S
Alonso
J C
JC
Lanka
E
E
Saenger
W
W
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D001412:Bacillus subtilis / Q000187:drug effects / Q000254:growth & development; D002455:Cell Division / Q000187:drug effects; D004265:DNA Helicases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004268:DNA-Binding Proteins; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development; D005419:Flavonoids / Q000737:chemistry / Q000494:pharmacology*; D007700:Kinetics; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013329:Structure-Activity Relationship; D015534:Trans-Activators
D004268:DNA-Binding Proteins; D005419:Flavonoids; D009841:Oligonucleotides; D011506:Proteins; D015534:Trans-Activators; C045264:replication initiator protein; C040015:myricetin; D000251:Adenosine Triphosphatases; D004265:DNA Helicases