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Nucleic Acids Research, 1985, Vol. 13, No. 19 6847-6866
© 1985

Articles

The distribution and properties of RNA primed initiation sites of DNA synthesis at the replication origin of Escherichia coli chromosome

Yuji Kohara, Naoki Tohdoh*, Xiao-Wang Jiang and Tuneko Okazaki

Institute of Molecular Biology, Faculty of Science, Nagoya University Nagoya 464, Japan

Received July 29, 1985. Accepted September 9, 1985.

RNA-linked DNA molecules were obtained from E. coli dnaCts cells synchronously initiating a new round of chromosome replication. The deoxynucleotides at the transition from primer RNA to DNA were 32P-labeled, and their positions were located on the nucleotide sequence of 1.4 kb genomic region (position –906 to +493) including the oriC and its leftside flanking region. In the r-strand (the counterclockwise strand), many strong transition sites ware mapped in the left half portion of the oriC and a few weak sites in the left outside region. In the l-strand (the clockwise strand), no transition sites were found inside the oriC but many weak sites were found in the left outside region. The results support the initiation mechanism in which the first leading strand synthesis starts with the r-strand counterclockwise from the oriC that is followed by the l-strand synthesis on the displaced template strand on the left of oriC. Primer RNA molecules attached to the strong r-strand transition sites were only a few residues in length. Properties of the transition sites were discussed.

*Present address: Biology Division, National Cancer Center Research Institute, Tsukiji 5-1-1, Chuo-ku, Tokyo 104, Japan


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