Presence of Proteins Derived from the Vegetative Cell Membrane in the Dormant Spore Coat of <i>Bacillus subtilis</i>

Fujita, Yūkō; Yasuda, Yoko; Kozuka, Satoshi; Tochikubo, Kunio

doi:10.1111/j.1348-0421.1989.tb01987.x

Cited by 9 publications

(5 citation statements)

References 28 publications

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“…Despite the lack of conclusive morphological evidence for the presence of an outer spore membrane, there is functional and biochemical evidence to support the presence of such a structure in the mature spore. For instance, there is evidence that 11 spore coat proteins are related antigenically to membrane proteins from vegetative cells (Fujita et al. 1989; as cited in Henriques and Moran 2000).…”

Section: Outer Membranementioning

confidence: 99%

Bacterial spore structures and their protective role in biocide resistance

et al. 2012

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Summary The structure and chemical composition of bacterial spores differ considerably from those of vegetative cells. These differences largely account for the unique resistance properties of the spore to environmental stresses, including disinfectants and sterilants, resulting in the emergence of spore‐forming bacteria such as Clostridium difficile as major hospital pathogens. Although there has been considerable work investigating the mechanisms of action of many sporicidal biocides against Bacillus subtilis spores, there is far less information available for other species and particularly for various Clostridia. This paucity of information represents a major gap in our knowledge given the importance of Clostridia as human pathogens. This review considers the main spore structures, highlighting their relevance to spore resistance properties and detailing their chemical composition, with a particular emphasis on the differences between various spore formers. Such information will be vital for the rational design and development of novel sporicidal chemistries with enhanced activity in the future.

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Section: Outer Membranementioning

confidence: 99%

Bacterial spore structures and their protective role in biocide resistance

et al. 2012

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“…There is evidence for the outer forespore membrane remaining intact in spores of B. megaterium (67), and this issue must be resolved in B. subtilis. Fujita et al (32) demonstrated the presence of some vegetative cell membrane proteins in the coat, possibly indicating that at least some of the outer forespore membrane is still intact in the dormant spore.…”

Section: Spore Morphologymentioning

confidence: 99%

Bacillus subtilis Spore Coat

Driks

1999

Microbiol Mol Biol Rev

485

299

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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.

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“…Alternatively, the inner coat itself may function as the permeability barrier and pretreatment with oxalic acid or phenol at a high temperature may cause partial destruction of the inner coat layer and enable the dyes to reach the outer pericortex region because spores treated with oxalic acid or phenol became susceptible to lysozyme ( Table 1). Failure of the dyes to penetrate the cortex suggests the existence of a secondary permeability barrier, that is, an outer membrane, which has been demonstrated biochemically in the spores of Bacillus megaterium (CraftsLighty & Ellar, 1980), by the morphological observation of variant spores lacking an exosporium (Koshikawa et al, 1984) and by showing the presence of vegetative cell membrane derived proteins in the spore coat of B. subtilis (Fujita et al, 1989). However, the localization and the integration of an outer membrane remain unclear with B. subtilis spores.…”

Section: Discussionmentioning

confidence: 99%

Permeability of dormant spores of Bacillus subtilis to malachite green and crystal violet

Kozuka

Tochikubo

1991

Journal of General Microbiology

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The permeability of dormant spores of Baciffus subtifis to malachite green (MG) and crystal violet (CV) was examined by using potassium trichloro(q2-ethylene)platinm(II) (KTR) as an electron-opaque marker for the dyes. The spores were treated with the dyes and other substances at 30 "C for 30 rnin or at 80 "C for 5 min. When the spores were incubated in 50 mM-MG solution at 30 "C and in 50 mM-CV solution at 30 "C or 80 "C, many small electron-dense precipitates, which were chemical complexes of dyes and platinum, were seen, mainly around the boundary between the inner and outer coat regions. The spores treated under the above conditions were not stained. Treatment with 50 mM-MG alone or a mixture of 25 mM-oxalic acid and 50 mM-CV at 80 "C made the spores stainable and dye-TPt precipitates were observed mainly in the outer pericortex region. Pretreatment with 25 mMoxalic acid and 5 % (v/v) phenol at 80 "C followed by 50 mM-CV treatment at 30 "C gave the same results as above.It was considered from these results that the inner coat itself might function as the primary permeability barrier to MG and CV, and that a secondary barrier to the dyes might exist around the cortex region. IntroductionBacterial endospores are well known for their extreme resistance and dormancy. Previous studies related to the permeability of dormant spores to disinfectants (Sykes, 1970; Thomas & Russell, 1974), polymyxin B (Tochikubo et al., 1986), lysozyme (Gould & Hitchins, 1963;Gould et al., 1970) and germinants (Koshikawa et al., 1984;Vary, 1973;Watabe et al., 1974) indicate that a spore coat or an outer pericortex membrane may play an important role as a penetration barrier. Moreover, indirect evidence for the existence of a penetration barrier at or near the outer edge of the cortex has been obtained by electron microscopy of unfixed spores embedded in methacrylate (Rode et al., 1962). Recently, we showed by immunoelectron microscopy that the permeability barrier of dormant spores of Bacillus subtilis to gentamicin was present outside the cortex (Tochikubo et al., 1988). Dormant spores cannot be stained by conventional methods for staining vegetative bacterial cells. Many investigators have noticed that acid treatment before staining (Fitz-James et al., 1954;Lechtman et al., 1965;Maneval, 1941;Robinow, 1951) during staining (Dorner, 1922 ;Dutton, 1928 ;May, 1926) is necessary to stain bacterial spores. In this paper, we report how far basic dyes can penetrate into B. subtilis spores and why the spores can be stained by the method of Schaeffer & Fulton (1933), which is a simplification of a method proposed by Wirtz (1908). MethodsDormant spores. Dormant spores of B. subtilis PCI 2 19, prepared by growing the organism on nutrient agar at 37 "C for 5 d, were harvested and washed five times with sterile deionized water by centrifugation.Treatment of dormant spores with dyes and other chemicals. Dormant spores (about 2 x lo9 ml-l) were suspended in an aqueous solution containing 50 mM-crystal violet hydrochloride (CV ; pH 3-9) or 50 mMm...

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Presence of Proteins Derived from the Vegetative Cell Membrane in the Dormant Spore Coat of Bacillus subtilis

Abstract: To confirm the presence of the outer spore membrane in dormant spore

Cited by 9 publications

References 28 publications

Bacterial spore structures and their protective role in biocide resistance

Bacterial spore structures and their protective role in biocide resistance

Bacillus subtilis Spore Coat

Permeability of dormant spores of Bacillus subtilis to malachite green and crystal violet

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