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Journal of Bacteriology, July 2001, p. 4115-4126, Vol. 183, No. 14
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.14.4115-4126.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Constitutive Septal Murein Synthesis in Escherichia coli with Impaired Activity of the Morphogenetic Proteins RodA and Penicillin-Binding Protein 2

Miguel A. de Pedro,^1,* William D. Donachie,² Joachim-Volker Höltje,³ and Heinz Schwarz³

Centro de Biología Molecular "Severo Ochoa," Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Campus de Cantoblanco, 28049 Madrid, Spain¹; Institute of Cell and Molecular Biology, The University of Edinburgh, Edinburgh EH9 3JR, Scotland²; and Max-Planck Institut für Entwitcklungsbiologie, D-7400 Tübingen, Germany³

Received 20 February 2001/Accepted 27 April 2001

The pattern of peptidoglycan (murein) segregation in cells of Escherichia coli with impaired activity of the morphogenetic proteins penicillin-binding protein 2 and RodA has been investigated by the D-cysteine-biotin immunolabeling technique (M. A. de Pedro, J. C. Quintela, J.-V. Höltje, and H. Schwarz, J. Bacteriol. 179:2823-2834, 1997). Inactivation of these proteins either by amdinocillin treatment or by mutations in the corresponding genes, pbpA and rodA, respectively, leads to the generation of round, osmotically stable cells. In normal rod-shaped cells, new murein precursors are incorporated all over the lateral wall in a diffuse manner, being mixed up homogeneously with preexisting material, except during septation, when strictly localized murein synthesis occurs. In contrast, in rounded cells, incorporation of new precursors is apparently a zonal process, localized at positions at which division had previously taken place. Consequently, there is no mixing of new and old murein. Old murein is preserved for long periods of time in large, well-defined areas. We propose that the observed patterns are the result of a failure to switch off septal murein synthesis at the end of septation events. Furthermore, the segregation results confirm that round cells of rodA mutants do divide in alternate, perpendicular planes as previously proposed (K. J. Begg and W. D. Donachie, J. Bacteriol. 180:2564-2567, 1998).

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^* Corresponding author. Mailing address: Centro de Biología Molecular "Severo Ochoa," CSIC-UAM, Facultad de Ciencias UAM, Campus de Cantoblanco, 28049 Madrid, Spain. Phone: (34) 913978083. Fax: (34) 913978087. E-mail: madepedro{at}cbm.uam.es.

This paper is dedicated to J. de la Rosa for the many years of hard work that she has dedicated to our group at Centro de Biología Molecular "Severo Ochoa."

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Journal of Bacteriology, July 2001, p. 4115-4126, Vol. 183, No. 14
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.14.4115-4126.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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