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Title: Coordinated Assembly of the Bacillus anthracis Coat and Exosporium during Bacterial Spore Outer Layer Formation

Abstract

Bacterial spores produced by the Bacillales are composed of concentric shells, each of which contributes to spore function. Spores from all species possess a cortex and coat, but spores from many species possess additional outer layers. The outermost layer of Bacillus anthracis spores, the exosporium, is separated from the coat by a gap known as the interspace. Exosporium and interspace assembly remains largely mysterious. As a result, we have a poor understanding of the overarching mechanisms driving the assembly of one of the most ubiquitous cell types in nature. To elucidate the mechanisms directing exosporium assembly, we generated strains bearing mutations in candidate exosporium-controlling genes and analyzed the effect on exosporium formation. Biochemical and cell biological analyses argue that CotE directs the assembly of CotO into the spore and that CotO might be located at or close to the interior side of the cap. Taken together with data showing that CotE and CotO interact directly in vitro, we propose a model in which CotE and CotO are important components of a protein interaction network that connects the exosporium to the forespore during cap formation and exosporium elongation. Our data also suggest that the cap interferes with coat assembly at onemore » pole of the spore, altering the pattern of coat deposition compared to the model organism Bacillus subtilis. We propose that the difference in coat assembly patterns between these two species is due to an inherent flexibility in coat assembly, which may facilitate the evolution of spore outer layer complexity.« less

Authors:
 [1];  [1];  [1];  [2];  [1];  [1];  [3];  [3];  [3];  [2];  [1];  [4];  [5]
+ Show Author Affiliations
  1. Loyola Univ. Chicago, Maywood, IL (United States). Dept. of Microbiology and Immunology
  2. Univ. of Missouri, Columbia, MO (United States). Dept. of Veterinary Pathobiology and Bond Life Sciences Center
  3. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Biosciences and Biotechnology Div.
  4. Univ. of Texas Houston, Houston, TX (United States)
  5. Harvard Univ., Cambridge, MA (United States)
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE National Nuclear Security Administration (NNSA)
OSTI Identifier:
1560106
Report Number(s):
LLNL-JRNL-774758
Journal ID: ISSN 2150-7511; 967108
Grant/Contract Number:  
AC52-07NA27344
Resource Type:
Accepted Manuscript
Journal Name:
mBio (Online)
Additional Journal Information:
Journal Name: mBio (Online); Journal Volume: 9; Journal Issue: 6; Journal ID: ISSN 2150-7511
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Boone, Tyler J., Mallozzi, Michael, Nelson, Alex, Thompson, Brian, Khemmani, Mark, Lehmann, Dörte, Dunkle, Alexis, Hoeprich, Paul, Rasley, Amy, Stewart, George, Driks, Adam, Koehler, Theresa M., and Losick, Richard. Coordinated Assembly of the Bacillus anthracis Coat and Exosporium during Bacterial Spore Outer Layer Formation. United States: N. p., 2018. Web. doi:10.1128/mBio.01166-18.
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Boone, Tyler J., Mallozzi, Michael, Nelson, Alex, Thompson, Brian, Khemmani, Mark, Lehmann, Dörte, Dunkle, Alexis, Hoeprich, Paul, Rasley, Amy, Stewart, George, Driks, Adam, Koehler, Theresa M., & Losick, Richard. Coordinated Assembly of the Bacillus anthracis Coat and Exosporium during Bacterial Spore Outer Layer Formation. United States. doi:10.1128/mBio.01166-18.
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Boone, Tyler J., Mallozzi, Michael, Nelson, Alex, Thompson, Brian, Khemmani, Mark, Lehmann, Dörte, Dunkle, Alexis, Hoeprich, Paul, Rasley, Amy, Stewart, George, Driks, Adam, Koehler, Theresa M., and Losick, Richard. Tue . "Coordinated Assembly of the Bacillus anthracis Coat and Exosporium during Bacterial Spore Outer Layer Formation". United States. doi:10.1128/mBio.01166-18. https://www.osti.gov/servlets/purl/1560106.
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@article{osti_1560106,
title = {Coordinated Assembly of the Bacillus anthracis Coat and Exosporium during Bacterial Spore Outer Layer Formation},
author = {Boone, Tyler J. and Mallozzi, Michael and Nelson, Alex and Thompson, Brian and Khemmani, Mark and Lehmann, Dörte and Dunkle, Alexis and Hoeprich, Paul and Rasley, Amy and Stewart, George and Driks, Adam and Koehler, Theresa M. and Losick, Richard},
abstractNote = {Bacterial spores produced by the Bacillales are composed of concentric shells, each of which contributes to spore function. Spores from all species possess a cortex and coat, but spores from many species possess additional outer layers. The outermost layer of Bacillus anthracis spores, the exosporium, is separated from the coat by a gap known as the interspace. Exosporium and interspace assembly remains largely mysterious. As a result, we have a poor understanding of the overarching mechanisms driving the assembly of one of the most ubiquitous cell types in nature. To elucidate the mechanisms directing exosporium assembly, we generated strains bearing mutations in candidate exosporium-controlling genes and analyzed the effect on exosporium formation. Biochemical and cell biological analyses argue that CotE directs the assembly of CotO into the spore and that CotO might be located at or close to the interior side of the cap. Taken together with data showing that CotE and CotO interact directly in vitro, we propose a model in which CotE and CotO are important components of a protein interaction network that connects the exosporium to the forespore during cap formation and exosporium elongation. Our data also suggest that the cap interferes with coat assembly at one pole of the spore, altering the pattern of coat deposition compared to the model organism Bacillus subtilis. We propose that the difference in coat assembly patterns between these two species is due to an inherent flexibility in coat assembly, which may facilitate the evolution of spore outer layer complexity.},
doi = {10.1128/mBio.01166-18},
journal = {mBio (Online)},
number = 6,
volume = 9,
place = {United States},
year = {2018},
month = {11}
}
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DOI:  10.1128/mBio.01166-18
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Figures / Tables:

Figure 1 Figure 1: Mutations in cotO result in the loss of the exosporium. Wild-type (A, G and H), MGM76 (cotOΩpMGM3; B, I and J), TJB124 (cotO; C, K and L), TJB130 (cotO, eag1ΩpTJB10; D, M and N), TJB116 (Sterne 34F2 [pMK4-cotO]; E) and TJB239 (Sterne $Δ$cot$Y$; F) spores were analyzed bymore » thin-section TEM (A-F) and immunofluorescence microscopy (G-N). Arrows indicate the exosporium, while arrowheads indicate the coat. In A, the cortex (Cx) and interspace (IS) are also indicated. Images are representative of at least 80 spores visualized. The presence of the exosporium was also assessed by immunofluorescence microscopy, staining with anti- BclA antibodies (G-N). Images are representative of more than 100 spores visualized in three separate spore preparations.« less
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