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Proteomic Studies of Bacillus anthracis Reveal In Vitro CO2-Modulation and Expression During Infection of Extracellular Proteases

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The Challenge of Highly Pathogenic Microorganisms

Abstract

A comparative proteomic study of secretomes of virulent and avirulent Bacillus anthracis strains in various culturing conditions, including those encountered in the host (high CO2/bicarbonate), enabled identification of approximately 70 proteins representing collectively more than 99% of the secretome. In-vivo expression of 50 proteins was established by 2-dimension Western-analysis using anti B. anthracis immune sera. Many of the abundant proteins harbor features characteristic of virulence determinants and exhibit different patterns of expression. In minimal medium, virulent and avirulent B. anthracis strains manifest similar protein signatures and the metalloprotease NprA, (previously suggested to act in the context of a starvation-induced mechanism), represents 90% of the total secretome. Under high CO2/bicarbonate, NprA is repressed (possibly by a mechanism which preserves toxin integrity), while other proteins, including the bacterial toxins, are induced. One of the immunogens observed to be induced under high CO2-tension, was HtrA. We investigated the phenotype associated with disruption of HtrA by biochemical and proteomic approaches. The HtrA- bacteria are severely affected in their ability to respond to stress and fail to secrete the most abundant extracellular protease NprA. Most surprisingly, HtrA- cells do not possess the characteristic S-layer. This unique phenotype may have important implications for the role of HtrA in manifestation of B. anthracis virulence. Furthermore, the data show that distinct CO2/bicarbonate responsive chromosome-and plasmid-encoded regulatory factors modulate the secretion of potential novel virulence factors, most of which are associated with extracellular proteolytic activities.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, P.O. Box 19, Ness-Ziona, 74100, Israel

    Theodor Chitlaru, Orit Gat, Galia Zaide, Haim Grosfeld, Izhak Inbar, Sharon Ehrlich, Moshe Leitner, Ofer Cohen & Avigdor Shafferman

Authors
  1. Theodor Chitlaru
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  2. Orit Gat
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  3. Galia Zaide
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  4. Haim Grosfeld
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  5. Izhak Inbar
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  6. Sharon Ehrlich
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  7. Moshe Leitner
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  8. Ofer Cohen
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  9. Avigdor Shafferman
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Corresponding author

Correspondence to Theodor Chitlaru .

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Editors and Affiliations

  1. Israel Institute for Biological Research, Ness-Ziona, 74100, Israel

    Avigdor Shafferman

  2. Israel Institute for Biological Research, P.O. Box 19, Ness-Ziona, 74100, Israel

    Arie Ordentlich

  3. Israel Institute for Biological Research, P.O. Box 19, Ness-Ziona, 74100, Israel

    Baruch Velan

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Chitlaru, T. et al. (2010). Proteomic Studies of Bacillus anthracis Reveal In Vitro CO2-Modulation and Expression During Infection of Extracellular Proteases. In: Shafferman, A., Ordentlich, A., Velan, B. (eds) The Challenge of Highly Pathogenic Microorganisms. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9054-6_2

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