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Nucleic Acids Research, 1983, Vol. 11, No. 18 6381-6397
© 1983

MOLECULAR BIOLOGY

Structural analysis of RNA molecules involved in plasmld copy number control

Joseph Tamm and Barry Polisky*

Program in Molecular, Cell, and Developmental Biology, Department of Biology, Indiana University Bloomington, IN 47405, USA

*To whom correspondence should be addressed

Received June 17, 1983. Revised August 30, 1983. Accepted August 30, 1983.

DNA replication of the plasmid ColE1 and its relatives is controlled mainly by a small plasmid-encoded transcript known as RNA1. We have studied the confor mation of RNA1 molecules from two related but compatible plasmids, ColEl and RSF1030, using T1 ribonu clease, S2nuclease, cobra venom nuclease, and diethyl pyrocarbonate modification as probes for secondary structure. Both RNA1 molecules contain three double-stranded stems, three single-stranded loops, and an exposed 5' tail.All loops in both molecules show similar sensitivities to the probes used, as do stems 1 and 3. The region comprising stem 2of each RNA1 molecule contains the most sequence differences as well as the most structural changes of any region in the two molecules. The structure of the RNA1 molecule encoded by a recessive high copy number mutant of ColEl was also investigated. Structural alterations involving the first stem and loop of the molecule are proposed to be responsible for the inability of the mutant RNA1 to function in vivo.


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