Replication of M‐13 DNA in Plasmolysed Escherichia coli Cells: Structure of a Replicative Intermediate with Restricted Binding of Intercalating Dyes

Barbara E. KESSLER‐LIEBSCHER, Walter L. STAUDENBAUER, Peter Hans HOFSCHNEIDER

Research output: Contribution to journalArticlepeer-review

Abstract

DNA molecules with restricted binding of intercalating dyes are observed as replicative intermediates during the replication of bacteriophage M‐13 duplex DNA in a cellular system in vitro prepared by plasmolysis of M‐13‐am5‐infected Escherichia coli cells. Restriction of dye binding is abolished by heating the DNA to 80 °C, but can be recovered by slow cooling of the heat‐treated DNA. Radioactive pulse‐label incorporated by these molecules is found exclusively in elongated viral strands of more than one genome length. In the electron microscope this DNA fraction is seen to contain a significant number of duplex DNA rings with two single‐stranded tails protruding from the same region of the ring. It is proposed that these structures arise by branch migration during the isolation of replicating molecules containing only one single‐stranded tail. The topological constraint in these molecules is most likely caused by base‐pairing between partially complementary regions of the two single‐stranded tails.

Original languageEnglish
Pages (from-to)233-239
Number of pages7
JournalEuropean Journal of Biochemistry
Volume84
Issue number1
DOIs
StatePublished - Mar 1978
Externally publishedYes

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