3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination

ABSTRACT Bacillus cereus, a Gram-positive aerobic bacterium commonly found in soil, food, and water, forms endospores that can withstand harsh environmental conditions. The endospores are encased in a protective spore coat consisting of multiple layers of proteins, among which, CotE serves as a cruc...

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Main Authors: Dukwon Lee, Yeongjin Baek, Migak Park, Doyeon Kim, Kyumi Byun, Jaekyung Hyun, Nam-Chul Ha
Format: Article
Language:English
Published: American Society for Microbiology 2025-04-01
Series:mBio
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Online Access:https://journals.asm.org/doi/10.1128/mbio.02472-24
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author Dukwon Lee
Yeongjin Baek
Migak Park
Doyeon Kim
Kyumi Byun
Jaekyung Hyun
Nam-Chul Ha
author_facet Dukwon Lee
Yeongjin Baek
Migak Park
Doyeon Kim
Kyumi Byun
Jaekyung Hyun
Nam-Chul Ha
author_sort Dukwon Lee
collection DOAJ
description ABSTRACT Bacillus cereus, a Gram-positive aerobic bacterium commonly found in soil, food, and water, forms endospores that can withstand harsh environmental conditions. The endospores are encased in a protective spore coat consisting of multiple layers of proteins, among which, CotE serves as a crucial morphogenetic protein within the outer coat. In this study, we observed that the homotrimeric CotE protein underwent further oligomerization induced by Ca2+ and was subsequently dissociated by dipicolinic acid, a compound released from the spore core during germination. Through cryo-electron microscopy and tomography analyses of the Ca2+-induced CotE oligomer, combined with structural predictions and biochemical studies, we propose a three-dimensional meshwork organization facilitated by tryptophan-based interactions between CotE trimers. The resulting meshwork was organized in a defective diamond-like tetrahedral configuration. These insights enhance our understanding of how CotE contributes to endospore morphogenesis and germination through the rapid disassembly of these layers.IMPORTANCEBacterial endospores are highly resilient structures that allow bacteria to survive extreme environmental conditions, making them a significant concern in food safety and healthcare. The protein CotE plays a critical role in forming the protective outer coat of these endospores. Our research uncovers the three-dimensional meshwork architecture of CotE and reveals how it contributes to the structural integrity and rapid disassembly of endospores during germination. By understanding CotE’s unique 3D structure and its interaction with other molecules, we gain valuable insights into how bacterial endospores are formed and how they can be effectively targeted for sterilization. This work not only advances our fundamental knowledge of bacterial endospore biology but also has potential applications in developing new strategies to combat bacterial contamination and improve sterilization techniques in the food and healthcare industries.
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spelling doaj-art-484c5c87cb074d81a5af6735d0fa9b292025-08-20T02:07:59ZengAmerican Society for MicrobiologymBio2150-75112025-04-0116410.1128/mbio.02472-243D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germinationDukwon Lee0Yeongjin Baek1Migak Park2Doyeon Kim3Kyumi Byun4Jaekyung Hyun5Nam-Chul Ha6Department of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, Center for Food and Bioconvergence, Interdisciplinary Programs in Agricultural Genomics, CALS, Seoul National University, Seoul, South KoreaDepartment of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, Center for Food and Bioconvergence, Interdisciplinary Programs in Agricultural Genomics, CALS, Seoul National University, Seoul, South KoreaDepartment of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, Center for Food and Bioconvergence, Interdisciplinary Programs in Agricultural Genomics, CALS, Seoul National University, Seoul, South KoreaDepartment of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, Center for Food and Bioconvergence, Interdisciplinary Programs in Agricultural Genomics, CALS, Seoul National University, Seoul, South KoreaSchool of Biological Sciences, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South KoreaSchool of Pharmacy, Sungkyunkwan University, Suwon, South KoreaDepartment of Agricultural Biotechnology, Research Institute of Agriculture and Life Sciences, Center for Food and Bioconvergence, Interdisciplinary Programs in Agricultural Genomics, CALS, Seoul National University, Seoul, South KoreaABSTRACT Bacillus cereus, a Gram-positive aerobic bacterium commonly found in soil, food, and water, forms endospores that can withstand harsh environmental conditions. The endospores are encased in a protective spore coat consisting of multiple layers of proteins, among which, CotE serves as a crucial morphogenetic protein within the outer coat. In this study, we observed that the homotrimeric CotE protein underwent further oligomerization induced by Ca2+ and was subsequently dissociated by dipicolinic acid, a compound released from the spore core during germination. Through cryo-electron microscopy and tomography analyses of the Ca2+-induced CotE oligomer, combined with structural predictions and biochemical studies, we propose a three-dimensional meshwork organization facilitated by tryptophan-based interactions between CotE trimers. The resulting meshwork was organized in a defective diamond-like tetrahedral configuration. These insights enhance our understanding of how CotE contributes to endospore morphogenesis and germination through the rapid disassembly of these layers.IMPORTANCEBacterial endospores are highly resilient structures that allow bacteria to survive extreme environmental conditions, making them a significant concern in food safety and healthcare. The protein CotE plays a critical role in forming the protective outer coat of these endospores. Our research uncovers the three-dimensional meshwork architecture of CotE and reveals how it contributes to the structural integrity and rapid disassembly of endospores during germination. By understanding CotE’s unique 3D structure and its interaction with other molecules, we gain valuable insights into how bacterial endospores are formed and how they can be effectively targeted for sterilization. This work not only advances our fundamental knowledge of bacterial endospore biology but also has potential applications in developing new strategies to combat bacterial contamination and improve sterilization techniques in the food and healthcare industries.https://journals.asm.org/doi/10.1128/mbio.02472-24CotEendosporeBacillus cereuscryo-electron microscopycryo-electron tomography
spellingShingle Dukwon Lee
Yeongjin Baek
Migak Park
Doyeon Kim
Kyumi Byun
Jaekyung Hyun
Nam-Chul Ha
3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination
mBio
CotE
endospore
Bacillus cereus
cryo-electron microscopy
cryo-electron tomography
title 3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination
title_full 3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination
title_fullStr 3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination
title_full_unstemmed 3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination
title_short 3D meshwork architecture of the outer coat protein CotE: implications for bacterial endospore sporulation and germination
title_sort 3d meshwork architecture of the outer coat protein cote implications for bacterial endospore sporulation and germination
topic CotE
endospore
Bacillus cereus
cryo-electron microscopy
cryo-electron tomography
url https://journals.asm.org/doi/10.1128/mbio.02472-24
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