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J. Biochem, 1988, Vol. 104, No. 3 333-336
© 1988 Japanese Biochemical Society

research-article

Phospholipid Modulates In Vitro Replication of Autonomous Replicating Sequence from Human Cells1

Hayato Umekawa*, Keiko Tamiya-Koizumi*, Eishi Takagi*, Hiroyoshi Ariga**, Kiyohide Kojima* and Shonen Yoshida*,2

*Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine Showa-ku, Nagoya, Aichi 466
**lnstitute of Medical Science, The University of Tokyo Minato-ku, Tokyo 108

2To whom correspondence should be addressed.

A cloned plasmid, pmyc(H-K), containing sequences derived from human c-myc gene replicated in vitro in Raji nuclear extract in a semiconservative manner. Using this system, it was found that phosphatidylinositol and cardiolipin strongly inhibited the replication of pmyc(H-K) in vitro, whereas other phospholipids, i.e., phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine, phosphatidic acid, and sphingomyelin, had no appreciable effect. The concentrations of phosphatidylinositol and cardiolipin producing 50% inhibition of the replication were 4.6 and 5.4 µM, respectively. Phosphatidylinositol and cardiolipin inhibited the relaxation of pmyc(H-K) supercoiled DNA, but showed little or weaker effects on DNA polymerase {alpha} and topoisomerase II in Raji nuclear extract. These results suggest that phosphatidylinositol and cardiolipin antagonize the replication of pmyc(H-K) in vitro, through, at least in part, the interaction with topoisomerase I.

1This work was supported in part by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.


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