Summary
Hemimethylated DNA occurs briefly in E.coli after passage of the replication fork and correlates well with the inhibition of the functioning of a number of genes involved in DNA replication including initiation of DNA synthesis at oriC and pBR322 origins of replication and transcription of the initiator DnaA gene. Previously, we found that inhibition of initiation in vitro at oriC on a hemimethylated DNA template relies on strong affinity between (a) membrane component(s) and the hemimethylated oriC sequence. Here, we show that this affinity is also shared by hemimethylated DNA fragments carrying the dnaA promoter and the RNA II promoter region which furnishes primers for pBR322 DNA replication. Thus, sequestration of hemimethylated DNA by the cell membrane may represent a common mechanism for the inhibition of replication and transcription at these and other sites on newly replicated DNA. We discuss these findings with reference to the control of interinitiation times of DNA replication. Also, we further characterize the membrane fraction by identifying a 24kDa protein responsible for the specific affinity for hemimethylated oriC DNA and describe hemimethylated oriC — membrane binding sensitivity to the sulfhydryl (-SH) group blocking agent, N-ethylmaleimide.
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© 1992 Springer-Verlag Berlin Heidelberg
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Hughes, P., Malki, A., Herrick, J., Kern, R., Kohiyama, M. (1992). Further Characterization of a DNA Replication Inhibitor from the Cell Membrane of Escherichia coli . In: Hughes, P., Fanning, E., Kohiyama, M. (eds) DNA Replication: The Regulatory Mechanisms. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-76988-7_4
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DOI: https://doi.org/10.1007/978-3-642-76988-7_4
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-76990-0
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