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Journal of Bacteriology, January 2004, p. 164-178, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.164-178.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Formation and Composition of the Bacillus anthracis Endospore{dagger}

Hongbin Liu,1,{ddagger} Nicholas H. Bergman,2,3,{ddagger} Brendan Thomason,2,{ddagger} Shamira Shallom,4 Alyson Hazen,4 Joseph Crossno,2 David A. Rasko,4 Jacques Ravel,4 Timothy D. Read,4,§ Scott N. Peterson,4,5,¶ John Yates III,1,¶ and Philip C. Hanna2,¶*

Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037,1 Department of Microbiology & Immunology,2 Bioinformatics Program, University of Michigan Medical School, Ann Arbor, Michigan 48109,3 Department of Microbial Genomics, The Institute for Genomic Research, Rockville, Maryland 20850,4 Department of Biochemistry and Molecular Biology, The George Washington University, Washington, D.C. 200105

Received 26 August 2003/ Accepted 30 September 2003

The endospores of Bacillus anthracis are the infectious particles of anthrax. Spores are dormant bacterial morphotypes able to withstand harsh environments for decades, which contributes to their ability to be formulated and dispersed as a biological weapon. We monitored gene expression in B. anthracis during growth and sporulation using full genome DNA microarrays and matched the results against a comprehensive analysis of the mature anthrax spore proteome. A large portion (~36%) of the B. anthracis genome is regulated in a growth phase-dependent manner, and this regulation is marked by five distinct waves of gene expression as cells proceed from exponential growth through sporulation. The identities of more than 750 proteins present in the spore were determined by multidimensional chromatography and tandem mass spectrometry. Comparison of data sets revealed that while the genes responsible for assembly and maturation of the spore are tightly regulated in discrete stages, many of the components ultimately found in the spore are expressed throughout and even before sporulation, suggesting that gene expression during sporulation may be mainly related to the physical construction of the spore, rather than synthesis of eventual spore content. The spore also contains an assortment of specialized, but not obviously related, metabolic and protective proteins. These findings contribute to our understanding of spore formation and function and will be useful in the detection, prevention, and early treatment of anthrax. This study also highlights the complementary nature of genomic and proteomic analyses and the benefits of combining these approaches in a single study.

* Corresponding author. Mailing address: University of Michigan Medical School, 5641 Med Sci II, Ann Arbor, MI 48109-0620. Phone: (734) 615-3706. Fax: (734) 764-3562. E-mail: pchanna{at}umich.edu.

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

{ddagger} H.L., N.H.B., and B.T. contributed equally to this study.

§ Present address: Biodefense Research Directorate, Naval Medical Research Center, Silver Spring, MD 20910.

S.N.P., J.Y., and P.C.H. contributed equally to this study.

Journal of Bacteriology, January 2004, p. 164-178, Vol. 186, No. 1
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.1.164-178.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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