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Molecular and Cellular Biology, February 2005, p. 888-895, Vol. 25, No. 3
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.3.888-895.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Mechanisms of Transcription-Replication Collisions in Bacteria

Ekaterina V. Mirkin and Sergei M. Mirkin*

Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, Illinois

Received 22 June 2004/ Returned for modification 22 October 2004/ Accepted 1 November 2004

While collisions between replication and transcription in bacteria are deemed inevitable, the fine details of the interplay between the two machineries are poorly understood. In this study, we evaluate the effects of transcription on the replication fork progression in vivo, by using electrophoresis analysis of replication intermediates. Studying Escherichia coli plasmids, which carry constitutive or inducible promoters in different orientations relative to the replication origin, we show that the mutual orientation of the two processes determines their mode of interaction. Replication elongation appears not to be affected by transcription proceeding in the codirectional orientation. Head-on transcription, by contrast, leads to severe inhibition of the replication fork progression. Furthermore, we evaluate the mechanism of this inhibition by limiting the area of direct contact between the two machineries. We observe that replication pausing zones coincide exactly with transcribed DNA segments. We conclude, therefore, that the replication fork is most likely attenuated upon direct physical interaction with the head-on transcription machinery.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, 900 S. Ashland Ave., Chicago, IL 60607. Phone: (312) 996-9610. Fax: (312) 413-0353. E-mail: mirkin{at}uic.edu.

Molecular and Cellular Biology, February 2005, p. 888-895, Vol. 25, No. 3
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.3.888-895.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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