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Journal of Bacteriology, November 2009, p. 6501-6512, Vol. 191, No. 21
0021-9193/09/$08.00+0     doi:10.1128/JB.00767-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Cell Wall Hydrolases Affect Germination, Vegetative Growth, and Sporulation in Streptomyces coelicolor{triangledown} ,{dagger}

Henry J. Haiser, Mary R. Yousef, and Marie A. Elliot*

Department of Biology and Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1280 Main Street West, Hamilton, Ontario L8S 4K1, Canada

Received 12 June 2009/ Accepted 20 August 2009

Peptidoglycan is a major cell wall constituent of gram-positive bacteria. It is a dynamic macromolecule that is actively remodeled to enable cell growth and differentiation through a tightly choreographed interplay of hydrolytic and biosynthetic enzyme activities. The filamentous bacterium Streptomyces coelicolor has a complex life cycle that likely requires considerable cell wall remodeling to enable both extension of vegetative hyphae and formation of differentiated cell types. In silico analysis of the S. coelicolor genome enabled identification of 56 candidate cell wall hydrolase genes. We found that seven of these genes shared a highly conserved 5' untranslated region and were expressed during both vegetative growth and sporulation; four of these genes were selected for more extensive biochemical and biological characterization. The proteins encoded by these genes, termed RpfA, SwlA, SwlB, and SwlC, were confirmed to be hydrolytic enzymes, as they could efficiently cleave S. coelicolor cell walls. Phenotypic analyses revealed that these enzymes are important throughout development; deletion of each hydrolase gene resulted in a mutant strain that was heat sensitive, defective in spore formation, and either altered in vegetative growth or delayed in spore germination. Our results indicate that these enzymes play key roles at multiple stages in the growth and development of S. coelicolor, highlighting both the lack of redundancy in hydrolase activity and the importance of cell wall remodeling in the S. coelicolor life cycle.

* Corresponding author. Mailing address: Department of Biology, McMaster University, 1280 Main Street West, Hamilton, ON L8S 4K1, Canada. Phone: (905) 525-9140, ext. 24225. Fax: (905) 522-6066. E-mail: melliot{at}mcmaster.ca

{triangledown} Published ahead of print on 28 August 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, November 2009, p. 6501-6512, Vol. 191, No. 21
0021-9193/09/$08.00+0     doi:10.1128/JB.00767-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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