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Journal of Bacteriology, March 2006, p. 1721-1732, Vol. 188, No. 5
0021-9193/06/$08.00+0     doi:10.1128/JB.188.5.1721-1732.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Localization of the Bacillus subtilis murB Gene within the dcw Cluster Is Important for Growth and Sporulation

Gonçalo Real and Adriano O. Henriques*

Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras Codex, Portugal

Received 22 August 2005/ Accepted 12 October 2005

The Bacillus subtilis murB gene, encoding UDP-N-acetylenolpyruvoylglucosamine reductase, a key enzyme in the peptidoglycan (PG) biosynthetic pathway, is embedded in the dcw (for "division and cell wall") cluster immediately upstream of divIB. Previous attempts to inactivate murB were unsuccessful, suggesting its essentiality. Here we show that the cell morphology, growth rate, and resistance to cell wall-active antibiotics of murB conditional mutants is a function of the expression level of murB. In one mutant, in which murB was insertionally inactivated in a merodiploid bearing a second xylose-inducible PxylA-murB allele, DivIB levels were reduced and a normal growth rate was achieved only if MurB levels were threefold that of the wild-type strain. However, expression of an extra copy of divIB restored normal growth at wild-type levels of MurB. In contrast, DivIB levels were normal in a second mutant containing an in-frame deletion of murB ({Delta}murB) in the presence of the PxylA-murB gene. Furthermore, this strain grew normally with wild-type levels of MurB. During sporulation, the levels of MurB were highest at the time of synthesis of the spore cortex PG. Interestingly, the {Delta}murB PxylA-murB mutant did not sporulate efficiently even at high concentrations of inducer. Since high levels of inducer did not interfere with sporulation of a murB+PxylA-murB strain, it appears that ectopic expression of murB fails to support efficient sporulation. These data suggest that coordinate expression of divIB and murB is important for growth and sporulation. The genetic context of the murB gene within the dcw cluster is unique to the Bacillus group and, taken together with our data, suggests that in these species it contributes to the optimal expression of cell division and PG biosynthetic functions during both vegetative growth and spore development.

* Corresponding author. Mailing address: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901 Oeiras Codex, Portugal. Phone: 351-21-4469521. Fax: 351-21-4411277. E-mail: aoh{at}itqb.unl.pt.

Journal of Bacteriology, March 2006, p. 1721-1732, Vol. 188, No. 5
0021-9193/06/$08.00+0     doi:10.1128/JB.188.5.1721-1732.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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