Phospholipid modulates in vitro replication of autonomous replicating sequence from human cells.
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 microM, 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.
Umekawa
H
H
Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Aichi.
Tamiya-Koizumi
K
K
Takagi
E
E
Ariga
H
H
Kojima
K
K
Yoshida
S
S
D004261:DNA Replication; D004264:DNA Topoisomerases, Type I / Q000032:analysis; D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D006801:Humans; D010743:Phospholipids / Q000502:physiology*; D010957:Plasmids; D011519:Proto-Oncogenes
D010743:Phospholipids; D004259:DNA-Directed DNA Polymerase; D004264:DNA Topoisomerases, Type I