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Nucleic Acids Research, 1988, Vol. 16, No. 19 9127-9145
© 1988

Articles

Functional analysis of the leading strand replication origin of plasmid pUB110 in Bacillus subtilis

Juan C. Alonso*, Heinrich Leonhardt and Carola A. Stiege

Max-Planck-Institut fur Molekulare Genetik Ihnestrasse 73, D-1000 Berlin 33, FRG

*To whom correspondence should be addressed

Received July 22, 1988. Accepted August 30, 1988.

Supercoiled plasmid DNA is the substrate for initiation of pUB110 replication, and - by inference - for binding of its initiator protein (RepU) to the plasmid replication origin (oriU) in vivo. No hairpin structure is required for RepU-oriU recognition. RepH (the pC194 replication initiation protein) failed to initiate replication in trans at oriU.

The nucleotides that determine the specificity of the replication initiation process are located within oriU but termination is unefficient. Therefore the segment that forms the full recognition signal for termination is probably located 3' of the oriU recognition sequence. Two overlapping domains, one for initiation and one required for termination, compose the leading strand replication origin of plasmid pUB110.


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