Localization of Proteins to Different Layers and Regions of<i>Bacillus subtilis</i>Spore Coats

Imamura, Daisuke; Kuwana, Ritsuko; Takamatsu, Hiromu; Watabe, Kazuhito

doi:10.1128/jb.01103-09

Cited by 59 publications

(55 citation statements)

References 45 publications

(72 reference statements)

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“…We detected major differences in the genes involved in spore coat structure and its biosynthesis between B. megaterium and B. subtilis strain 168 (see Table S3 in the supplemental material). More than half of the B. subtilis genes involved in spore coat formation are missing and could not be identified in either B. megaterium strain, which suggests that B. megaterium possesses a different spore coat structure than B. subtilis (23).…”

Section: Resultsmentioning

confidence: 91%

Genome Sequences of the Biotechnologically Important Bacillus megaterium Strains QM B1551 and DSM319

et al. 2011

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Bacillus megaterium is deep-rooted in the Bacillus phylogeny, making it an evolutionarily key species and of particular importance in understanding genome evolution, dynamics, and plasticity in the bacilli. B. megaterium is a commercially available, nonpathogenic host for the biotechnological production of several substances, including vitamin B 12 , penicillin acylase, and amylases. Here, we report the analysis of the first complete genome sequences of two important B. megaterium strains, the plasmidless strain DSM319 and QM B1551, which harbors seven indigenous plasmids. The 5.1-Mbp chromosome carries approximately 5,300 genes, while QM B1551 plasmids represent a combined 417 kb and 523 genes, one of the largest plasmid arrays sequenced in a single bacterial strain. We have documented extensive gene transfer between the plasmids and the chromosome. Each strain carries roughly 300 strain-specific chromosomal genes that account for differences in their experimentally confirmed phenotypes. B. megaterium is able to synthesize vitamin B 12 through an oxygen-independent adenosylcobalamin pathway, which together with other key energetic and metabolic pathways has now been fully reconstructed. Other novel genes include a second ftsZ gene, which may be responsible for the large cell size of members of this species, as well as genes for gas vesicles, a second ␤-galactosidase gene, and most but not all of the genes needed for genetic competence. Comprehensive analyses of the global Bacillus gene pool showed that only an asymmetric region around the origin of replication was syntenic across the genus. This appears to be a characteristic feature of the Bacillus spp. genome architecture and may be key to their sporulating lifestyle.

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Section: Resultsmentioning

confidence: 91%

Genome Sequences of the Biotechnologically Important Bacillus megaterium Strains QM B1551 and DSM319

et al. 2011

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“…Importantly, a group of proteins affected by deletion of cotE may be associated with the coat. For instance, the BC_5391 protein, absent in ⌬cotE spores formed at 37°C, is an orthologue of the B. subtilis coat protein GerQ, which is most likely localized in the inner coat (17,64). Although deletion of gerQ (ywdL) in B. cereus leads to exosporium fragility, this effect could be due to an abnormal assembly of the underlying coat (12).…”

Section: Discussionmentioning

confidence: 99%

“…Studies in the model organism Bacillus subtilis have evidenced the role of a group of so-called morphogenetic proteins in directing the assembly of the coat components (10). In B. subtilis, the coat is differentiated into an inner layer, an outer layer, and a more external structure called the crust (16)(17)(18). Dedicated morphogenetic proteins govern the assembly of the various coat substructures.…”

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Sporulation Temperature Reveals a Requirement for CotE in the Assembly of both the Coat and Exosporium Layers of Bacillus cereus Spores

Bressuire-Isoard

Bornard²,

Henriques

et al. 2016

Appl Environ Microbiol

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The Bacillus cereus spore surface layers consist of a coat surrounded by an exosporium. We investigated the interplay between the sporulation temperature and the CotE morphogenetic protein in the assembly of the surface layers of B. cereus ATCC 14579 spores and on the resulting spore properties. The cotE deletion affects the coat and exosporium composition of the spores formed both at the suboptimal temperature of 20°C and at the optimal growth temperature of 37°C. Transmission electron microscopy revealed that ⌬cotE spores had a fragmented and detached exosporium when formed at 37°C. However, when produced at 20°C, ⌬cotE spores showed defects in both coat and exosporium attachment and were susceptible to lysozyme and mutanolysin. Thus, CotE has a role in the assembly of both the coat and exosporium, which is more important during sporulation at 20°C. CotE was more represented in extracts from spores formed at 20°C than at 37°C, suggesting that increased synthesis of the protein is required to maintain proper assembly of spore surface layers at the former temperature. ⌬cotE spores formed at either sporulation temperature were impaired in inosine-triggered germination and resistance to UV-C and H 2 O 2 and were less hydrophobic than wild-type (WT) spores but had a higher resistance to wet heat. While underscoring the role of CotE in the assembly of B. cereus spore surface layers, our study also suggests a contribution of the protein to functional properties of additional spore structures. Moreover, it also suggests a complex relationship between the function of a spore morphogenetic protein and environmental factors such as the temperature during spore formation. Bacterial endospores are formed in a wide range of ecological niches in soil, as well as in the gastrointestinal tract of invertebrate and vertebrate animals, and in both natural and anthropized environments (1). Physical and chemical conditions prevailing in such niches play a major role in triggering sporulation and in determining the final properties of the resulting spores (2). Laboratory experiments demonstrate the major influence of environment, in particular of temperature, on the efficiency and yield of sporulation, and in spore resistance to wet heat, UV, high hydrostatic pressure, and preservatives or spore response to germinants (3; reviewed in references 1 and 4). Spore resistance and functional properties result from the assembly of several protective structures: cortex, coat, and exosporium. The spore peptidoglycan cortex, a structure common to all endospores, is a major factor in the resistance of spores to heat (1, 4). The cortex is surrounded by a proteinaceous coat, and in organisms such as Bacillus anthracis or Bacillus cereus the coat is further enveloped by an exosporium, a "balloon-like" structure consisting of a paracrystalline basal layer and an external hair-like nap formed mainly by the collagen-like glycoprotein BclA (5-11). While the coat contributes to protection against peptidoglycan-breaking enzymes, UV light, and oxidative...

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“…cotVWXYZ is a cluster of genes transcribed at the late stage of sporulation in mother cells from the promoters P cotVWX , P cotX , and P cotYZ (27). Because CotW and CotZ are localized to the spore crust (4,16), it is conceivable that the genes in the cluster are involved in spore crust formation. In order to identify genes important for spore crust formation, we monitored CgeA-GFP in the cotV, cotW, cotX, cotY, and cotZ mutant strains (Fig.…”

Section: Resultsmentioning

confidence: 99%

Proteins Involved in Formation of the Outermost Layer of Bacillus subtilis Spores

et al. 2011

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To investigate the outermost structure of the Bacillus subtilis spore, we analyzed the accessibility of antibodies to proteins on spores of B. subtilis. Anti-green fluorescent protein (GFP) antibodies efficiently accessed GFP fused to CgeA or CotZ, which were previously assigned to the outermost layer termed the spore crust. However, anti-GFP antibodies did not bind to spores of strains expressing GFP fused to 14 outer coat, inner coat, or cortex proteins. Anti-CgeA antibodies bound to spores of wild-type and CgeA-GFP strains but not cgeA mutant spores. These results suggest that the spore crust covers the spore coat and is the externally exposed, outermost layer of the B. subtilis spore. We found that CotZ was essential for the spore crust to surround the spore but not for spore coat formation, indicating that CotZ plays a critical role in spore crust formation. In addition, we found that CotY-GFP was exposed on the surface of the spore, suggesting that CotY is an additional component of the spore crust. Moreover, the localization of CotY-GFP around the spore depended on CotZ, and CotY and CotZ depended on each other for spore assembly. Furthermore, a disruption of cotW affected the assembly of CotV-GFP, and a disruption of cotX affected the assembly of both CotV-GFP and CgeA-GFP. These results suggest that cgeA and genes in the cotVWXYZ cluster are involved in spore crust formation.

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Localization of Proteins to Different Layers and Regions ofBacillus subtilisSpore Coats

Cited by 59 publications

References 45 publications

Genome Sequences of the Biotechnologically Important Bacillus megaterium Strains QM B1551 and DSM319

Genome Sequences of the Biotechnologically Important Bacillus megaterium Strains QM B1551 and DSM319

Sporulation Temperature Reveals a Requirement for CotE in the Assembly of both the Coat and Exosporium Layers of Bacillus cereus Spores

Proteins Involved in Formation of the Outermost Layer of Bacillus subtilis Spores

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