scholarly journals Localization of Proteins to Different Layers and Regions of Bacillus subtilis Spore Coats

2009 ◽  
Vol 192 (2) ◽  
pp. 518-524 ◽  
Author(s):  
Daisuke Imamura ◽  
Ritsuko Kuwana ◽  
Hiromu Takamatsu ◽  
Kazuhito Watabe
Keyword(s):  
Bacillus Subtilis ◽  
Mother Cell ◽  
Fluorescent Proteins ◽  
Bacterial Spores ◽  
Spore Coat ◽  
Subtilis Spore ◽  
Proximal Pole ◽  
Outer Coat ◽  
Outermost Layer ◽  
Bacillus Subtilis Spore

ABSTRACT Bacterial spores are encased in a multilayered proteinaceous shell known as the coat. In Bacillus subtilis, over 50 proteins are involved in spore coat assembly but the locations of these proteins in the spore coat are poorly understood. Here, we describe methods to estimate the positions of protein fusions to fluorescent proteins in the spore coat by using fluorescence microscopy. Our investigation suggested that CotD, CotF, CotT, GerQ, YaaH, YeeK, YmaG, YsnD, and YxeE are present in the inner coat and that CotA, CotB, CotC, and YtxO reside in the outer coat. In addition, CotZ and CgeA appeared in the outermost layer of the spore coat and were more abundant at the mother cell proximal pole of the forespore, whereas CotA and CotC were more abundant at the mother cell distal pole of the forespore. These polar localizations were observed both in sporangia prior to the release of the forespore from the mother cell and in mature spores after release. Moreover, CotB was observed at the middle of the spore as a ring- or spiral-like structure. Formation of this structure required cotG expression. Thus, we conclude not only that the spore coat is a multilayered assembly but also that it exhibits uneven spatial distribution of particular proteins.

scholarly journals Assembly of Multiple CotC Forms into the Bacillus subtilis Spore Coat

2004 ◽  
Vol 186 (4) ◽  
pp. 1129-1135 ◽  
Author(s):  
Rachele Isticato ◽  
Giovanni Esposito ◽  
Rita Zilhão ◽  
Sofia Nolasco ◽  
Giuseppina Cangiano ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Posttranslational Modifications ◽  
Mother Cell ◽  
Spore Coat ◽  
Spore Surface ◽  
Subtilis Spore ◽  
Cell Compartment ◽  
Bacillus Subtilis Spore ◽  
Molecular Masses

ABSTRACT We report evidence that the CotC polypeptide, a previously identified component of the Bacillus subtilis spore coat, is assembled into at least four distinct forms. Two of these, having molecular masses of 12 and 21 kDa, appeared 8 h after the onset of sporulation and were probably assembled on the forming spore immediately after their synthesis, since no accumulation of either of them was detected in the mother cell compartment, where their synthesis occurs. The other two components, 12.5 and 30 kDa, were generated 2 h later and were probably the products of posttranslational modifications of the two early forms occurring directly on the coat surface during spore maturation. None of the CotC forms was found either on the spore coat or in the mother cell compartment of a cotH mutant. This indicates that CotH serves a dual role of stabilizing the early forms of CotC and promoting the assembly of both early and late forms on the spore surface.

scholarly journals The Timing of cotE Expression Affects Bacillus subtilis Spore Coat Morphology but Not Lysozyme Resistance

2006 ◽  
Vol 189 (6) ◽  
pp. 2401-2410 ◽  
Author(s):  
Teresa Costa ◽  
Mónica Serrano ◽  
Leif Steil ◽  
Uwe Völker ◽  
Charles P. Moran ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Mother Cell ◽  
Spore Coat ◽  
Outer Coat ◽  
Bacillus Subtilis Spore ◽  
Spore Resistance ◽  
Early Expression ◽  
Cell Gene Expression ◽  
Coat Layer

ABSTRACT The synthesis of structural components and morphogenetic factors required for the assembly of the Bacillus subtilis spore coat is governed by a mother cell-specific transcriptional cascade. The first two temporal classes of gene expression, which involve RNA polymerase sigma σE factor and the ancillary regulators GerR and SpoIIID, are deployed prior to engulfment of the prespore by the mother cell. The two last classes rely on σK, whose activation follows engulfment completion, and GerE. The cotE gene codes for a morphogenetic protein essential for the assembly of the outer coat layer and spore resistance to lysozyme. cotE is expressed first from a σE-dependent promoter and, in a second stage, from a promoter that additionally requires SpoIIID and that remains active under σK control. CotE localizes prior to engulfment completion close to the surface of the developing spore, but formation of the outer coat is a late, σK-controlled event. We have transplanted cotE to progressively later classes of mother cell gene expression. This created an early class of mutants in which cotE is expressed prior to engulfment completion and a late class in which expression of cotE follows the complete engulfment of the prespore. Mutants of the early class assemble a nearly normal outer coat structure, whereas mutants of the late class do not. Hence, the early expression of CotE is essential for outer coat assembly. Surprisingly, however, all mutants were fully resistant to lysozyme. The results suggest that CotE has genetically separable functions in spore resistance to lysozyme and spore outer coat assembly.

scholarly journals CotC-CotU Heterodimerization during Assembly of the Bacillus subtilis Spore Coat

2007 ◽  
Vol 190 (4) ◽  
pp. 1267-1275 ◽  
Author(s):  
Rachele Isticato ◽  
Assunta Pelosi ◽  
Rita Zilhão ◽  
Loredana Baccigalupi ◽  
Adriano O. Henriques ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Mother Cell ◽  
Spore Coat ◽  
Spore Surface ◽  
Subtilis Spore ◽  
Cell Compartment ◽  
Sporulating Cell ◽  
Bacillus Subtilis Spore ◽  
Cell Chamber

ABSTRACT We report evidence that CotC and CotU, two previously identified components of the Bacillus subtilis spore coat, are produced concurrently in the mother cell chamber of the sporulating cell under the control of σK and GerE and immediately assembled around the forming spore. In the coat, the two proteins interact to form a coat component of 23 kDa. The CotU-CotC interaction was not detected in two heterologous hosts, suggesting that it occurs only in B. subtilis. Monomeric forms of both CotU and CotC failed to be assembled at the surface of the developing spore and accumulated in the mother cell compartment of cells mutant for cotE. In contrast, neither CotU nor CotC accumulated in the mother cell compartment of cells mutant for cotH. These results suggest that CotH is required to protect both CotU and CotC in the mother cell compartment of the sporangium and that CotE is needed to allow their assembly and subsequent interaction at the spore surface.

scholarly journals Laccase and Its Mutant Displayed on the Bacillus subtilis Spore Coat for Oxidation of Phenolic Compounds in Organic Solvents

Catalysts ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 606
Author(s):  
Silu Sheng ◽  
Edgardo T. Farinas
Keyword(s):  
Bacillus Subtilis ◽  
Organic Solvents ◽  
Environmental Changes ◽  
Sinapic Acid ◽  
Product Yield ◽  
Spore Coat ◽  
Subtilis Spore ◽  
Bacillus Subtilis Spore ◽  
Cota Laccase ◽  
Inert Surface

Enzymes displayed on the Bacillus subtilis spore coat have several features that are useful for biocatalysis. The enzyme is preimmobilized on an inert surface of the spore coat, which is due to the natural sporulation process. As a result, protein stability can be increased, and they are resistant to environmental changes. Next, they would not lyse under extreme conditions, such as in organic solvents. Furthermore, they can be easily removed from the reaction solution and reused. The laboratory evolved CotA laccase variant T480A-CotA was used to oxidize the following phenolic substrates: (+)-catechin, (−)-epicatechin, and sinapic acid. The kinetic parameters were determined and T480A-CotA had a greater Vmax/Km than wt-CotA for all substrates. The Vmax/Km for T480A-CotA was 4.1, 5.6, and 1.4-fold greater than wt-CotA for (+)-catechin, (−)-epicatechin, and sinapic acid, respectively. The activity of wt-CotA and T480A-CotA was measured at different concentrations from 0–70% in organic solvents (dimethyl sulfoxide, ethanol, methanol, and acetonitrile). The Vmax for T480A-CotA was observed to be greater than the wt-CotA in all organic solvents. Finally, the T480A-CotA was recycled 7 times over a 23-h period and up to 60% activity for (+)-catechin remained. The product yield was up to 3.1-fold greater than the wild-type.

Display of Bombyx mori Nucleopolyhedrovirus GP64 on the Bacillus subtilis Spore Coat

2011 ◽  
Vol 62 (5) ◽  
pp. 1368-1373 ◽  
Author(s):  
Guohui Li ◽  
Qi Tang ◽  
Huiqing Chen ◽  
Qin Yao ◽  
Degang Ning ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Bombyx Mori ◽  
Spore Coat ◽  
Subtilis Spore ◽  
Bombyx Mori Nucleopolyhedrovirus ◽  
Bacillus Subtilis Spore

scholarly journals Temporal and spatial regulation of protein cross-linking by the pre-assembled substrates of a Bacillus subtilis spore coat transglutaminase

PLoS Genetics ◽  
2019 ◽  
Vol 15 (4) ◽  
pp. e1007912 ◽  
Author(s):  
Catarina G. Fernandes ◽  
Diogo Martins ◽  
Guillem Hernandez ◽  
Ana L. Sousa ◽  
Carolina Freitas ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Cross Linking ◽  
Spore Coat ◽  
Subtilis Spore ◽  
Spatial Regulation ◽  
Bacillus Subtilis Spore ◽  
Temporal And Spatial ◽  
Protein Cross Linking

Regulation of the Transcription of a Cluster of Bacillus subtilis Spore Coat Genes

1994 ◽  
Vol 240 (5) ◽  
pp. 405-415 ◽  
Author(s):  
Jianke Zhang ◽  
Hiroshi Ichikawa ◽  
Richard Halberg ◽  
Lee Kroos ◽  
Arthur I. Aronson
Keyword(s):  
Bacillus Subtilis ◽  
Spore Coat ◽  
Subtilis Spore ◽  
Bacillus Subtilis Spore

scholarly journals Bacillus subtilis Spore Coat

1999 ◽  
Vol 63 (1) ◽  
pp. 1-20 ◽  
Author(s):  
Adam Driks
Keyword(s):  
Bacillus Subtilis ◽  
Posttranslational Processing ◽  
Spore Coat ◽  
Coat Proteins ◽  
Spore Surface ◽  
Subtilis Spore ◽  
Cell Type ◽  
Dormant Cell ◽  
Bacillus Subtilis Spore ◽  
Specialized Program

SUMMARY In response to starvation, bacilli and clostridia undergo a specialized program of development that results in the production of a highly resistant dormant cell type known as the spore. A proteinacious shell, called the coat, encases the spore and plays a major role in spore survival. The coat is composed of over 25 polypeptide species, organized into several morphologically distinct layers. The mechanisms that guide coat assembly have been largely unknown until recently. We now know that proper formation of the coat relies on the genetic program that guides the synthesis of spore components during development as well as on morphogenetic proteins dedicated to coat assembly. Over 20 structural and morphogenetic genes have been cloned. In this review, we consider the contributions of the known coat and morphogenetic proteins to coat function and assembly. We present a model that describes how morphogenetic proteins direct coat assembly to the specific subcellular site of the nascent spore surface and how they establish the coat layers. We also discuss the importance of posttranslational processing of coat proteins in coat morphogenesis. Finally, we review some of the major outstanding questions in the field.

scholarly journals Erratum. Bacillus subtilis spore coat assembly requires cotH gene expression.

1996 ◽  
Vol 178 (21) ◽  
pp. 6407-6407 ◽  
Author(s):  
G Naclerio ◽  
L Baccigalupi ◽  
R Zilhao ◽  
M De Felice ◽  
E Ricca
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Spore Coat ◽  
Subtilis Spore ◽  
Bacillus Subtilis Spore
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