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Expression of the Escherichia coli ftsZ gene: trials and tribulations of gene fusion studies

Published online by Cambridge University Press:  14 April 2009

Aline Robin  and
Richard D'Ari
Aline Robin*
Affiliation:
Institut Jacques Monod, C.N.R.S., Université Paris 7, 2, place Jussieu, 75251 Paris Cedex 05, France
Richard D'Ari
Affiliation:
Institut Jacques Monod, C.N.R.S., Université Paris 7, 2, place Jussieu, 75251 Paris Cedex 05, France
*
* Corresponding author.
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The ftsZ gene of Escherichia coli, which codes for an essential cell division protein, is subjected to multiple regulation, as shown in part with studies using an ftsZ::lacZ operon fusion located on phage λJFLIOO. Using this same fusion, we sought to isolate regulatory mutants overexpressing ftsZ by selecting mutants able to grow on lactose. One Lac+ mutant was obtained which overexpressed the ftsZ::lacZ fusion 70-fold. The mutation responsible for the overexpression lies in a new gene, cot, located near 56 min on the E. coli genetic map. The cot mutation probably affects the transcription of a chromosomal open reading frame, 0RF1, lying downstream of the bioA gene and adjacent to the ftzZ::lacZ fusion of the λJFL100 prophage integrated at attλ. Using an ftsZ84(Ts) strain, in which there was a double selection for overexpression of both ftsZ::lacZ and ftsZ+, no Lac+Tr mutants were obtained from 3·6 × 1010 bacteria; the introduction of a mutL allele, increasing spontaneous base substitution mutation rates 75-fold, did not permit us to isolate such a mutant. We conclude that Lac+ftsZ-constitutive mutations cannot be obtained in λJFL100 lysogens by a single base substitution.

Type
Research Article
Information
Genetics Research , Volume 61 , Issue 1 , February 1993 , pp. 1 - 8
Copyright
Copyright © Cambridge University Press 1993

References

Aldea, M., Garrido, T., Pla, J. & Vicente, M. (1990). Division genes in Escherichia coli are expressed coordinately to cell septum requirements by gearbox promoters. EMBO Journal 9, 3787–3794.CrossRefGoogle ScholarPubMed
Barker, D. F. & Campbell, A. M. (1980). Use of bio-lac fusion strains to study regulation of biotin biosynthesis in Escherichia coli. Journal of Bacteriology 143, 789–800.CrossRefGoogle ScholarPubMed
Bi, E. & Lutkenhaus, J. (1991). FtsZ ring structure associated with division in Escherichia coli. Nature 354, 161–164.CrossRefGoogle ScholarPubMed
Campbell, A., Del, Campillo-Campbell A. & Barker, D. (1978). Repression of biotin biosynthesis in Escherichia coli during growth on biotin vitamers. Journal of Bacteriology 135, 90–98.CrossRefGoogle ScholarPubMed
Carson, M. J., Barondess, J. & Beckwith, J. (1991). The FtsQ protein of Escherichia coli: membrane topology, abundance, and cell division phenotypes due to overproduction and insertion mutations. Journal of Bacteriology 173, 2187–2195.CrossRefGoogle ScholarPubMed
Corton, J. C, Ward, J. E. & Lutkenhaus, J. (1987). Analysis of cell division gene ftsZ(sulB) from gram-negative and gram-positive bacteria. Journal of Bacteriology 169, 1–7.CrossRefGoogle ScholarPubMed
Dai, K. & Lutkenhaus, J. (1991). ftsZ is an essential cell division gene in Escherichia coli. Journal of Bacteriology 173, 3500–3506.CrossRefGoogle ScholarPubMed
Dessen, P., Fondrat, C, Valencia, C. & Mugnier, C. (1990). BISANCE: a French service for access to biomolecules. Computer Applications in Biosciences 6, 355–356.Google Scholar
Dewar, S. J., Kagan-Zur, V., Begg, K. J. & Donachie, W. D. (1989). Transcriptional regulation of cell division genes in Escherichia coli. Molecular Microbiology 3, 1371–1377.CrossRefGoogle ScholarPubMed
Gervais, F. G., Phoenix, P. & Drapeau, G. R. (1992). The rcsB gene, a positive regulator of colanic acid biosynthesis in Escherichia coli, is also an activator of ftsZ expression. Journal of Bacteriology 174, 3964–3971.CrossRefGoogle ScholarPubMed
Ghelardini, P., Lauri, P., Ruberti, I., Orlando, V. & Paolozzi, L. (1991). Synchronous division induced in Escherichia coli K12 by phage Mu: analysis of DNA topology and gene expression during the cell cycle. Research in Microbiology 142, 259–267.CrossRefGoogle ScholarPubMed
Glickman, B. W. & Radman, M. (1980). Escherichia coli mutator mutants deficient in methylation instructed DNA mismatch correction. Proceedings of the National Academy of Sciences, USA 11, 1063–1067.CrossRefGoogle Scholar
Huisman, O., Jacques, M., D'Ari, R. & Caro, L. (1983). Role of the sfi-dependent cell division regulation system in Escherichia coli. Journal of Bacteriology 153, 1072–1074.CrossRefGoogle Scholar
Kravchenko, V. V., Vassilenko, S. K. & Grachev, M. A. (1979). A rightward promoter to the left of the att site of A phage DNA: possible participant in site-specific recombination. Gene 7, 181–195.CrossRefGoogle Scholar
Lutkenhaus, J. (1990). Regulation of cell division in E.coli. Trends in Genetics 6, 22–25.CrossRefGoogle ScholarPubMed
Masters, M., Paterson, T., Popplewell, A. G., Owen-Hughes, T., Pringle, J. H. & Begg, K. J. (1989). The effect of DnaA protein levels and rate of initiation at oriC on transcription originating in the ftsQ and ftsA genes: in vivo experiments. Molecular & General Genetics 216, 475–483.CrossRefGoogle ScholarPubMed
Miller, J. H. (1972). Experiments in Molecular Genetics. Cold Spring Harbor Laboratory; Cold Spring Harbor, N.Y.Google Scholar
Mizusawa, S. & Ward, D. F. (1982). A bacteriophage lambda vector for cloning with BamHl and SaulA. Gene 20,317–322.CrossRefGoogle Scholar
Otsuka, A. J., Buoncristiani, M. R., Howard, P. K., Flamm, J., Johnson, C, Yamamoto, R., Uchida, K., Cook, C, Ruppert, J. & Matsuzaki, J. (1988). The Escherichia coli biotin biosynthetic enzyme sequences predicted from the nucleotide sequence of the bio operon. Journal of Biological Chemistry 263, 19577–19585.CrossRefGoogle ScholarPubMed
Pearson, W. R. & Lipman, D. J. (1988). Improved tools for biological sequence comparison. Proceedings of the National Academy of Sciences, USA 85, 2444–2448.CrossRefGoogle ScholarPubMed
Rasmussen, L. J., Meller, P. L. & Atlung, T. (1991). Carbon metabolism regulates expression of the pfl (pyruvate formate-lyase) gene in Escherichia coli. Journal of Bacteriology 173, 6390–6397.CrossRefGoogle ScholarPubMed
Robin, A., Joseleau-Petit, D. & D'Ari, R. (1990). Transcription of the ftsZ gene and cell division in Escherichia coli. Journal of Bacteriology 172, 1392–1399.CrossRefGoogle ScholarPubMed
Robinson, A. C, Begg, K. J. & Donachie, W. D. (1988). Mapping and characterization of mutants of the Escherichia coli cell division gene, ftsA. Molecular Microbiology 2, 581–588.CrossRefGoogle ScholarPubMed
Rosenveld, E. C, Calva, E., Burgess, R. R. & Szybalski, W. (1980). In vitro transcription from the b2 region of bacteriophage A. Virology 107, 476–487.CrossRefGoogle Scholar
Ruberti, I., Crescenzi, F., Paolozzi, L. & Ghelardini, P. (1991). A class of gyrB mutants, substantially unaffected in DNA topology, suppresses the Escherichia coli K12 ftsZu mutation. Molecular Microbiology 5, 1065–1072.CrossRefGoogle ScholarPubMed
Schwartz, M. (1987). The maltose regulon. In Escherichia coli and Salmonella typhimurium: cellular and molecular biology (ed. Neidhardt, F., Ingraham, J. L., Low, K. B., Magasanik, B., Schaechter, M. and Umbarger, H. E.), pp. 1482–1502. Washington, D.C.: American Society for Microbiology.Google Scholar
Silhavy, T. J. & Beckwith, J. R. (1985). Use of lac fusions for the study of biological problems. Microbiological Reviews 49, 398–418.CrossRefGoogle Scholar
Vicente, M., Kushner, S. R., Garrido, T. & Aldea, M. (1991). The role of the ‘gearbox’ in the transcription of essential genes. Molecular Microbiology 5, 2085–2091.CrossRefGoogle ScholarPubMed
Walker, J. R., Kovarik, J. S., Allen, J. S. & Gustafson, R. A. (1975). Regulation of bacterial cell division: temperature-sensitive mutants of Escherichia coli that are defective in septum formation. Journal of Bacteriology 123, 693–703.CrossRefGoogle ScholarPubMed
Wang, X., de Boer, P. A. J. & Rothfield, L. I. (1991). A factor that positively regulates cell division by activating transcription of the major cluster of essential cell division genes of Escherichia coli. EMBO Journal 10, 3363–3372.CrossRefGoogle ScholarPubMed
Ward, J. E. Jr & Lutkenhaus, J. (1985). Overproduction of FtsZ induces minicell formation in E.coli. Cell 42, 941–949.CrossRefGoogle ScholarPubMed