scholarly journals Characterization of exosporium layer variability of Clostridioides difficile spores in the epidemically relevant strain R20291

2020 ◽  
Author(s):  
Marjorie Pizarro-Guajardo ◽  
Paulina Calderón ◽  
Alba Romero-Rodriguez ◽  
Daniel Paredes-Sabja
Keyword(s):  
Spore Coat ◽  
Coat Proteins ◽  
Anaerobic Conditions ◽  
Host Interactions ◽  
Clostridioides Difficile ◽  
Transmission Electron ◽  
Antibiotic Associated Diarrhea ◽  
Biological Aspects ◽  
Spore Coat Proteins

AbstractClostridioides difficile is a Gram-positive anaerobic intestinal pathogenic bacterium and the causative agent of antibiotic-associated diarrhea and spores are the transmission vehicle of the disease. In C. difficile spores, the outermost exosporium layer is the first barrier of interaction with the host and should carry spore ligands involved in spore-host interactions. C. difficile forms two types of spores (i.e., thin and thick exosporium layers). In this communication, we contribute to understand several biological aspects of these two exosporium morphotypes. By transmission electron microscopy, we demonstrate that both exosporium morphotypes appear simultaneously during sporulation and that the laminations of the spore-coat are formed under anaerobic conditions. Nycodenz density-gradient allows enrichment of spores with a thick-exosporium layer morphotype and presence of polar appendage. Using translational fluorescent fusions with exosporium proteins BclA3, CdeA, CdeC and CdeM as well as with several spore coat proteins, we observed that expression intensity and distribution of SNAP-translational fusions in R20291 strain is highly heterogeneous. Electron micrographs demonstrate that multicopy expression of CdeC, but not CdeM, SNAP translational fusion, increases the abundance of the thick exosporium morphotype. Collectively, these results raise further questions on how these distinctive exosporium morphotypes are made during spore formation.

Analyse ultrastructurale et biochimique de mutants pléiotropes de Bacillus subtilis affectés dans le contrôle de la sporulation

1984 ◽  
Vol 30 (11) ◽  
pp. 1367-1376
Author(s):  
Joseph Zucca ◽  
Serge Renaudin
Keyword(s):  
Bacillus Subtilis ◽  
Pleiotropic Effect ◽  
Spore Coat ◽  
Coat Proteins ◽  
Extracellular Proteases ◽  
Transmission Electron ◽  
Mutant Strains ◽  
Spore Coat Proteins ◽  
Spore Ultrastructure ◽  
Spore Coat Protein

Derived from a Bacillus subtilis unstable mutant strains which hyperproduce extracellular proteases were found to contain mutations at loci scoC, scoD, or scoE; these were observed by transmission electron microscopy. In scoC and scoD strains, spores formed overproduced spore coat proteins and the spore ultrastructure was deeply altered. In scoE strains, stage III of the sporulation was blocked. The pleiotropic effect of scoC mutations was either positive or negative. In unstable strains, mutations at the scoC locus stimulated α-amylase and levane sucrase hyperproduction. In stable strains, the same mutations led to an absence of α-amylase synthesis but to a normal levane sucrase production. ScoD mutations only induced α-amylase hyperproduction, while scoE mutations allowed neither levane sucrase production nor spore coat protein synthesis.

scholarly journals Functional Characterization of Clostridium difficile Spore Coat Proteins

2013 ◽  
Vol 195 (7) ◽  
pp. 1492-1503 ◽  
Author(s):  
P. Permpoonpattana ◽  
J. Phetcharaburanin ◽  
A. Mikelsone ◽  
M. Dembek ◽  
S. Tan ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Functional Characterization ◽  
Spore Coat ◽  
Coat Proteins ◽  
Spore Coat Proteins

scholarly journals SpoVID Guides SafA to the Spore Coat inBacillus subtilis

2001 ◽  
Vol 183 (10) ◽  
pp. 3041-3049 ◽  
Author(s):  
Amanda J. Ozin ◽  
Craig S. Samford ◽  
Adriano O. Henriques ◽  
Charles P. Moran
Keyword(s):  
Direct Interaction ◽  
Spore Coat ◽  
Coat Proteins ◽  
Yeast Two Hybrid System ◽  
Spore Coat Proteins ◽  
Basement Layer ◽  
Two Hybrid ◽  
Spore Coat Protein

ABSTRACT Bacteria assemble complex structures by targeting proteins to specific subcellular locations. The protein coat that encasesBacillus subtilis spores is an example of a structure that requires coordinated targeting and assembly of more than 24 polypeptides. The earliest stages of coat assembly require the action of three morphogenetic proteins: SpoIVA, CotE, and SpoVID. In the first steps, a basement layer of SpoIVA forms around the surface of the forespore, guiding the subsequent positioning of a ring of CotE protein about 75 nm from the forespore surface. SpoVID localizes near the forespore membrane where it functions to maintain the integrity of the CotE ring and to anchor the nascent coat to the underlying spore structures. However, it is not known which spore coat proteins interact directly with SpoVID. In this study we examined the interaction between SpoVID and another spore coat protein, SafA, in vivo using the yeast two-hybrid system and in vitro. We found evidence that SpoVID and SafA directly interact and that SafA interacts with itself. Immunofluorescence microscopy showed that SafA localized around the forespore early during coat assembly and that this localization of SafA was dependent on SpoVID. Moreover, targeting of SafA to the forespore was also dependent on SpoIVA, as was targeting of SpoVID to the forespore. We suggest that the localization of SafA to the spore coat requires direct interaction with SpoVID.

Cloning and expression of a cDNA that comprises part of the gene coding for a spore coat protein of Dictyostelium discoideum

1984 ◽  
Vol 4 (11) ◽  
pp. 2273-2278
Author(s):  
B C Dowds ◽  
W F Loomis
Keyword(s):  
Dictyostelium Discoideum ◽  
Cdna Clone ◽  
Single Gene ◽  
Polyclonal Antiserum ◽  
Cloning And Expression ◽  
Spore Coat ◽  
Coat Proteins ◽  
Developmentally Regulated ◽  
Gene Coding ◽  
Spore Coat Proteins

The three major spore coat proteins of Dictyostelium discoideum are developmentally regulated, cell-type-specific proteins. They are packaged in prespore vesicles and then secreted to form the outer layer of spore coats. We have isolated a cDNA clone from the gene coding for one of these proteins, SP96, a glycoprotein of 96,000 daltons. We screened the cDNA bank by the method of hybrid select translation followed by immunoprecipitation of the translation products with SP96-specific polyclonal antiserum. We found that the gene was first transcribed into stable mRNA a few hours before the time of detection of SP96 synthesis and that the mRNA, like the protein, accumulated specifically in prespore cells and spores. SP96 constituted the same proportion of newly synthesized protein as the proportion of its message in polyadenylated RNA. SP96 appeared to be encoded by a single gene as judged by Southern blot analysis of digested genomic DNA hybridized to the cDNA clone.

scholarly journals Synthesis and Deposition of Spore Coat Proteins during Sporulation ofBacillus megaterium

1985 ◽  
Vol 29 (12) ◽  
pp. 1151-1162 ◽  
Author(s):  
Masayoshi Imagawa ◽  
Yuichi Oku ◽  
Hussein I. El-Belbasi ◽  
Mie Teraoka ◽  
Tsutomu Nishihara ◽  
...  
Keyword(s):  
Spore Coat ◽  
Coat Proteins ◽  
Spore Coat Proteins

scholarly journals The yabG gene of Bacillus subtilis encodes a sporulation specific protease which is involved in the processing of several spore coat proteins

2000 ◽  
Vol 192 (1) ◽  
pp. 33-38 ◽  
Author(s):  
Hiromu Takamatsu ◽  
Atsuo Imamura ◽  
Takeko Kodama ◽  
Kei Asai ◽  
Naotake Ogasawara ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Spore Coat ◽  
Coat Proteins ◽  
Specific Protease ◽  
Spore Coat Proteins

Phosphorylation of spore coat proteins of Dictyostelium discoideum

1983 ◽  
Vol 61 (9) ◽  
pp. 996-1001 ◽  
Author(s):  
Teshome Akalehiywot ◽  
Chi-Hung Siu
Keyword(s):  
Electrophoretic Mobility ◽  
Dictyostelium Discoideum ◽  
Cellular Proteins ◽  
Developmental Cycle ◽  
Spore Coat ◽  
Coat Proteins ◽  
Phosphoamino Acid ◽  
Molecular Weights ◽  
Phosphorylated Proteins ◽  
Spore Coat Proteins

Phosphorylation of cellular proteins was studied during development of Dictyostelium discoideum. In the second half of the developmental cycle, two heavily phosphorylated proteins were observed together with a limited number of minor phosphorylated proteins. The electrophoretic mobility of these two phosphoproteins corresponded to two of the major spore coat glycoproteins, with apparent molecular weights of 103 000 and 80 000. They were found to be externalized and associated with the spore coat during spore formation. Phosphoserine was the predominant phosphoamino acid in both cases. These two phosphoproteins thus serve as excellent markers for the differentiation of prespore cells in D. discoideum.

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