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Journal of Bacteriology, January 2006, p. 687-693, Vol. 188, No. 2
0021-9193/06/$08.00+0     doi:10.1128/JB.188.2.687-693.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Phenotype Microarray Profiling of Staphylococcus aureus menD and hemB Mutants with the Small-Colony-Variant Phenotype{dagger}

Christof von Eiff,1* Peter McNamara,2 Karsten Becker,1 Donna Bates,2 Xiang-He Lei,3 Michael Ziman,3 Barry R. Bochner,3 Georg Peters,1 and Richard A. Proctor2,4

Institute of Medical Microbiology, University of Muenster, Muenster, Germany,1 Department of Medical Microbiology & Immunology; University of Wisconsin Medical School, Madison, Wisconsin,2 Biolog, Inc., Hayward, California,3 Department of Medicine, University of Wisconsin Medical School, Madison, Wisconsin4

Received 29 July 2005/ Accepted 14 October 2005

Standard biochemical tests have revealed that hemin and menadione auxotrophic Staphylococcus aureus small-colony variants (SCVs) exhibit multiple phenotypic changes. To provide a more complete analysis of the SCV phenotype, two genetically defined mutants with a stable SCV phenotype were comprehensively tested. These mutants, generated via mutations in menD or hemB that yielded menadione and hemin auxotrophs, were subjected to phenotype microarray (PM) analysis of over 1,500 phenotypes (including utilization of different carbon, nitrogen, phosphate, and sulfur sources; growth stimulation or inhibition by amino acids and other nutrients, osmolytes, and metabolic inhibitors; and susceptibility to antibiotics). Compared to parent strain COL, the hemB mutant was defective in utilization of a variety of carbon sources, including Krebs cycle intermediates and compounds that ultimately generate ATP via electron transport. The phenotype of the menD mutant was similar to that of the hemB mutant, but the defects in carbon metabolism were more pronounced than those seen with the hemB mutant. In both mutant strains, hexose phosphates and other carbohydrates that provide ATP in the absence of electron transport stimulated growth. Other phenotypes of SCV mutants, such as hypersensitivity to sodium selenite, sodium tellurite, and sodium nitrite, were also uncovered by the PM analysis. Key results of the PM analysis were confirmed in independent growth studies and by using Etest strips for susceptibility testing. PM technology is a new and efficient technology for assessing cellular phenotypes in S. aureus.

* Corresponding author. Mailing address: Institute of Medical Microbiology, University Hospital of Münster, Domagkstrasse 10, 48149 Münster, Germany. Phone: 49-251-835-5360. Fax: 49-251-835-5350. E-mail: eiffc{at}uni-muenster.de.

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

Journal of Bacteriology, January 2006, p. 687-693, Vol. 188, No. 2
0021-9193/06/$08.00+0     doi:10.1128/JB.188.2.687-693.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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