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Journal of Bacteriology, February 2004, p. 1205-1212, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1205-1212.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A Temperature-Sensitive Mutation in the dnaE Gene of Caulobacter crescentus That Prevents Initiation of DNA Replication but Not Ongoing Elongation of DNA

Teresa Lo,1 Ellena van Der Schalie,2 Theresa Werner,1 Yves V. Brun,1 and Neena Din2*

Department of Biology, Loyola College, Baltimore, Maryland 21210,2 Department of Biology, Indiana University, Bloomington, Indiana 474051

Received 31 January 2003/ Accepted 5 November 2003

A genetic screen for cell division cycle mutants of Caulobacter crescentus identified a temperature-sensitive DNA replication mutant. Genetic complementation experiments revealed a mutation within the dnaE gene, encoding the {alpha}-catalytic subunit of DNA polymerase III holoenzyme. Sequencing of the temperature-sensitive dnaE allele indicated a single base pair substitution resulting in a change from valine to glutamic acid within the C-terminal portion of the protein. This mutation lies in a region of the DnaE protein shown in Escherichia coli, to be important in interactions with other essential DNA replication proteins. Using DNA replication assays and fluorescence flow cytometry, we show that the observed block in DNA synthesis in the Caulobacter dnaE mutant strain occurs at the initiation stage of replication and that there is also a partial block of DNA elongation.

* Corresponding author. Mailing address: Loyola College, Department of Biology, 4501 North Charles St., Baltimore, MD 21210. Phone: (410) 617-2240. Fax: (410) 617-5682. E-mail: ndin1{at}loyola.edu.

Journal of Bacteriology, February 2004, p. 1205-1212, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1205-1212.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Galhardo, R. S., Rocha, R. P., Marques, M. V., Menck, C. F. M. (2005). An SOS-regulated operon involved in damage-inducible mutagenesis in Caulobacter crescentus. Nucleic Acids Res 33: 2603-2614 [Abstract] [Full Text]  


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