Fully methylated oriC with negative superhelicity forms an oriC-membrane complex before initiation of chromosome replication

Autor: Kazuo Nagai, S. Gayama, Makari Yamasaki, Takao Kataoka, Masaaki Wachi, J. Nakamura
Rok vydání: 1993
Předmět:
Zdroj: Biochemical and biophysical research communications. 194(3)
ISSN: 0006-291X
Popis: In an in vitro assay, the oriC DNA has been shown to bind to the outer membrane fraction only when it is hemimethylated (G.B.Ogden et al., Cell, 54, 127-135,1988). In this report, however, we demonstrated that a significant amount of the oriC DNA was recovered from the cells just before initiation with the oriC DNA being fully methylated. Formation of this preinitiation oriC-membrane complex and following initiation of chromosome replication were strongly inhibited by novobiocin, a DNA gyrase B subunit inhibitor, which reduced the superhelicity of the reporter plasmid in the cells. On the other hand, both reactions proceeded in the presence of nalidixic acid, a DNA gyrase A subunit inhibitor, which did not have the effect of reducing the superhelicity. These results suggest that the negative superhelicity of the DNA is required for preinitiation oriC-membrane complex formation and following initiation event of replication.
Databáze: OpenAIRE