BMQ_0737 encodes a novel protein crucial to the integrity of the outermost layers of<i>Bacillus megaterium</i>QM B1551 spores

Manetsberger, Julia; Hall, Elizabeth A. H.; Christie, Graham

doi:10.1111/1574-6968.12520

Cited by 4 publications

(7 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The construction of B. megaterium strains bearing C-terminal green fluorescent protein (GFP) fusions to SleL (BMQ_0021), BMQ_4051, BMQ_3035, and BMQ_0737 proteins has been described previously ( 10 ). Strains with GFP fusions to CotE (BMQ_4110) and CotW (BMQ_pBM60030) were prepared similarly.…”

Section: Methodsmentioning

confidence: 99%

Ellipsoid Localization Microscopy Infers the Size and Order of Protein Layers in Bacillus Spore Coats

Manetsberger

Manton

Erdélyi

et al. 2015

Biophysical Journal

Self Cite

View full text Add to dashboard Cite

Multilayered protein coats are crucial to the dormancy, robustness, and germination of bacterial spores. In Bacillus subtilis spores, the coat contains over 70 distinct proteins. Identifying which proteins reside in each layer may provide insight into their distinct functions. We present image analysis methods that determine the order and geometry of concentric protein layers by fitting a model description for a spheroidal fluorescent shell image to optical micrographs of spores incorporating fluorescent fusion proteins. The radius of a spherical protein shell can be determined with <10 nm error by fitting an equation to widefield fluorescence micrographs. Ellipsoidal shell axes can be fitted with comparable precision. The layer orders inferred for B. subtilis and B. megaterium are consistent with measurements in the literature. The aspect ratio of elongated spores and the tendency of some proteins to localize near their poles can be quantified, enabling measurement of structural anisotropy.

show abstract

Section: Methodsmentioning

confidence: 99%

Ellipsoid Localization Microscopy Infers the Size and Order of Protein Layers in Bacillus Spore Coats

Manetsberger

Manton

Erdélyi

et al. 2015

Biophysical Journal

Self Cite

View full text Add to dashboard Cite

show abstract

“…Planar inclusions were evident in the interspace region when spores were visualized by transmission electron microscopy. A novel morphogenetic spore coat protein, BMQ_0737, was found to be important for the proper assembly of the outer spore coat and exosporium layers of the B. megaterium QM B1551 spores (182). A pole-localized nap layer of strain QM B1551 is composed of the BclA1 protein, and this polar nap localization is independent of BxpB (183).…”

Section: Bacillus Megateriummentioning

confidence: 99%

The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host

Stewart

2015

Microbiol Mol Biol Rev

138

155

View full text Add to dashboard Cite

SUMMARYMuch of what we know regarding bacterial spore structure and function has been learned from studies of the genetically well-characterized bacteriumBacillus subtilis. Molecular aspects of spore structure, assembly, and function are well defined. However, certain bacteria produce spores with an outer spore layer, the exosporium, which is not present onB. subtilisspores. Our understanding of the composition and biological functions of the exosporium layer is much more limited than that of other aspects of the spore. Because the bacterial spore surface is important for the spore's interactions with the environment, as well as being the site of interaction of the spore with the host's innate immune system in the case of spore-forming bacterial pathogens, the exosporium is worthy of continued investigation. Recent exosporium studies have focused largely on members of theBacillus cereusfamily, principallyBacillus anthracisandBacillus cereus. Our understanding of the composition of the exosporium, the pathway of its assembly, and its role in spore biology is now coming into sharper focus. This review expands on a 2007 review of spore surface layers which provided an excellent conceptual framework of exosporium structure and function (A. O. Henriques and C. P. Moran, Jr., Annu Rev Microbiol61:555–588, 2007,http://dx.doi.org/10.1146/annurev.micro.61.080706.093224). That review began a process of considering outer spore layers as an integrated, multilayered structure rather than simply regarding the outer spore components as independent parts.

show abstract

“…Transformation was achieved using standard polyethylene glycol-mediated procedures, followed by recovery on hypertonic medium supplemented with antibiotics (33). Strains that had integrated pVLG6 plasmids to create gfp fusions were isolated, and the construct was verified by PCR, essentially as described previously (34). Spores were prepared by nutrient exhaustion in supplemented nutrient broth and subsequently purified by repeated rounds of centrifugation and resuspension of spore pellets in ice-cold water (33).…”

Section: Methodsmentioning

confidence: 99%

“…Microscopy. Fluorescence and thin-section transmission electron microscopy (TEM) analyses were conducted as described previously (34).…”

Section: Methodsmentioning

confidence: 99%

Orthologues of Bacillus subtilis Spore Crust Proteins Have a Structural Role in the Bacillus megaterium QM B1551 Spore Exosporium

Manetsberger

Ghosh

Hall

et al. 2018

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

The exosporium ofBacillus megateriumQM B1551 spores is morphologically distinct from exosporia observed for the spores of many other species. Previous work has demonstrated that unidentified genes carried on one of the large indigenous plasmids are required for the assembly of theBacillus megateriumexosporium. Here, we provide evidence that pBM600-encoded orthologues of theBacillus subtilisCotW and CotX proteins, which form the crust layer in spores of that species, are structural components of theBacillus megateriumQM B1551 spore exosporium. The introduction of plasmid-bornecotWand orthologouscotXgenes to the PV361 strain, which lacks all indigenous plasmids and produces spores that are devoid of an exosporium, results in the development of spores with a rudimentary exosporium-type structure. Additionally, purified recombinant CotW protein is shown to assemble at the air-water interface to form thin sheets of material, which is consistent with the idea that this protein may form a basal layer in theBacillus megateriumQM B1551 exosporium.IMPORTANCEWhen starved of nutrients, some bacterial species develop metabolically dormant spores that can persist in a viable state in the environment for several years. The outermost layers of spores are of particular interest since (i) these represent the primary site for interaction with the environment and (ii) the protein constituents may have biotechnological applications. The outermost layer, or exosporium, inBacillus megateriumQM B1551 spores is of interest, as it is morphologically distinct from the exosporia of spores of the pathogenicBacillus cereusfamily. In this work, we provide evidence that structurally important protein constituents of theBacillus megateriumexosporium are different from those in theBacillus cereusfamily. We also show that one of these proteins, when purified, can assemble to form sheets of exosporium-like material. This is significant, as it indicates that spore-forming bacteria employ different proteins and mechanisms of assembly to construct their external layers.

show abstract

BMQ_0737 encodes a novel protein crucial to the integrity of the outermost layers ofBacillus megateriumQM B1551 spores

Cited by 4 publications

References 22 publications

Ellipsoid Localization Microscopy Infers the Size and Order of Protein Layers in Bacillus Spore Coats

Ellipsoid Localization Microscopy Infers the Size and Order of Protein Layers in Bacillus Spore Coats

The Exosporium Layer of Bacterial Spores: a Connection to the Environment and the Infected Host

Orthologues of Bacillus subtilis Spore Crust Proteins Have a Structural Role in the Bacillus megaterium QM B1551 Spore Exosporium

Contact Info

Product

Resources

About