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Journal of Bacteriology, April 2007, p. 3187-3197, Vol. 189, No. 8
0021-9193/07/$08.00+0     doi:10.1128/JB.01846-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of Major Sporulation Proteins of Myxococcus xanthus Using a Proteomic Approach

John L. Dahl,^1,* Farah K. Tengra,^2, David Dutton,¹ Jinyuan Yan,² Tracy M. Andacht,³ Lia Coyne,¹ Veronica Windell,¹ and Anthony G. Garza²

School of Molecular Biosciences, Washington State University, Pullman, Washington 99164,¹ Department of Biology, Syracuse University, Syracuse, New York 13244,² Proteomics Resource Facility, Integrated Biotechnology Laboratories, University of Georgia, 458 Animal and Dairy Sciences Bldg., 425 River Road, Athens, Georgia 30602³

Received 8 December 2006/ Accepted 25 January 2007

Myxococcus xanthus is a soil-dwelling, gram-negative bacterium that during nutrient deprivation is capable of undergoing morphogenesis from a vegetative rod to a spherical, stress-resistant spore inside a domed-shaped, multicellular fruiting body. To identify proteins required for building stress-resistant M. xanthus spores, we compared the proteome of liquid-grown vegetative cells with the proteome of mature fruiting body spores. Two proteins, protein S and protein S1, were differentially expressed in spores, as has been reported previously. In addition, we identified three previously uncharacterized proteins that are differentially expressed in spores and that exhibit no homology to known proteins. The genes encoding these three novel major spore proteins (mspA, mspB, and mspC) were inactivated by insertion mutagenesis, and the development of the resulting mutant strains was characterized. All three mutants were capable of aggregating, but for two of the strains the resulting fruiting bodies remained flattened mounds of cells. The most pronounced structural defect of spores produced by all three mutants was an altered cortex layer. We found that mspA and mspB mutant spores were more sensitive specifically to heat and sodium dodecyl sulfate than wild-type spores, while mspC mutant spores were more sensitive to all stress treatments examined. Hence, the products of mspA, mspB, and mspC play significant roles in morphogenesis of M. xanthus spores and in the ability of spores to survive environmental stress.

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* Corresponding author. Mailing address: School of Molecular Biosciences, Washington State University, Abelson Hall, Room 301, Pullman, WA 99164. Phone: (509) 335-7719. Fax: (509) 335-1907. E-mail: johndahl{at}wsu.edu

Published ahead of print on 9 February 2007.

J.L.D. and F.K.T. contributed equally to this work.

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Journal of Bacteriology, April 2007, p. 3187-3197, Vol. 189, No. 8
0021-9193/07/$08.00+0     doi:10.1128/JB.01846-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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