The Bacillus subtilis Replication Terminator System Functions in Escherichia coli
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TL;DR
The Bacillus subtilis DNA terminators, IRI + IRII, were inserted into the Escherichia coli plasmid pACYC184 such that the IRI terminator would be in its active orientation with respect to the approaching unidirectionally moving replication fork.
Abstract
The Bacillus subtilis DNA terminators, IRI + IRII, were inserted into the Escherichia coli plasmid pACYC184 such that the IRI terminator would be in its active orientation with respect to the approaching unidirectionally moving replication fork. When this new plasmid was transferred into E. coli, harbouring an expression plasmid producing the B. subtilis terminator protein RTP, fork arrest was observed to occur at the position of the inserted terminator region. Thus, the B. subtilis replication terminator system can function in E. coli. It was shown that the B. subtilis system operated with approximately 30% of the efficiency of the E. coli system utilizing the R6K TerR2 DNA terminator and the E. coli Tus terminator protein. Assuming that RTP and Tus have quite different folded structures these results suggest that fork arrest in B. subtilis is not dependent upon a highly specific recognition and interaction between RTP positioned on the DNA terminator and a component(s) of the approaching replisome.