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Nucleic Acids Research, 1983, Vol. 11, No. 18 6531-6539
© 1983

MOLECULAR BIOLOGY

Flexbility in RNA priming of Okazald pieces at the E. coli replication fork

Kirk R. Thomas and Baldomero M. Olivera

Department of Biology, University of Utah Salt Lake City, UT 84112, USA

Received June 8, 1983. Revised August 22, 1983. Accepted August 22, 1983.

We present results which suggest considerable flexibility 1n the RNA priming of Okazakipieces at the E. coli replication fork. Using film lysates on cellophane discs, we have Identified RNA at the 5'ends of Okazakipieces. All four ribonucleotides are found to be present at the RNA-DNA junction 1f all four ribonucleoside triphosphates are used.However,1f only ATP,or ATP and GTP are used, then only 2'(3')AMP,or 2'(3')AMP and 2'(3')GMP are found at the RNA-DNA junction.A nearest neighbor analysis of RNA associated with Okazakipieces using a32P-CTP as a probe shows a similar dependence of nearest neighbor composition on the ribonucleoside triphosphate composition of the Incubation mixture.Thus, the nucleotide composition of the RNA primers at the ends of Okazaki pieces varies as a function of the ribonucleoside triphosphates available.


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