Stimulation of DNA polymerase alpha by hypergravity generated by centrifugal acceleration.
Gravity alteration is known to influence cell proliferation. Here we tested the effects of hypergravity on the action of DNA polymerase alpha, one of the DNA replication enzymes in eukaryotes. Hypergravity was produced by horizontal centrifugal acceleration with a hand-made rotator. The reaction rate of DNA polymerase alpha in centrifuge tubes increased along with the acceleration up to 4g, when a plateau was reached. In contrast, no stimulation was observed with primase, DNA polymerase epsilon, and the E. coli DNA polymerase I Klenow fragment. Kinetic analysis of DNA polymerase alpha reactions revealed that, under high gravity conditions, the K(m) value for template DNA decreased while the V(max) stayed constant. In contrast, the centrifugal acceleration did not affect the K(m) values for deoxyribonucleoside triphosphates. These results suggest that the hypergravity enhances the activity of DNA polymerase alpha by increasing the affinity of the enzyme for template DNA. Such enhancement was more prominent with a low concentration of DNA polymerase alpha under low ionic conditions.
Takemura
M
M
Laboratory of Cancer Cell Biology, Nagoya University School of Medicine, Nagoya 466-8550, Japan.
Yoshida
S
S
D000818:Animals; D002417:Cattle; D002498:Centrifugation / Q000295:instrumentation* / Q000379:methods*; D004256:DNA Polymerase I / Q000737:chemistry* / Q000378:metabolism*; D004257:DNA Polymerase II / Q000737:chemistry; D019915:DNA Primase / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D006112:Gravitation; D006367:HeLa Cells; D006801:Humans; D007477:Ions; D007700:Kinetics
D007477:Ions; D019915:DNA Primase; D004256:DNA Polymerase I; D004257:DNA Polymerase II