Localized insertion of new S-layer during growth of Bacillus stearothermophilus strains

Gruber, Karl Heinz; Sleytr, Uwe B.

doi:10.1007/bf00446749

Cited by 23 publications

(14 citation statements)

References 27 publications

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“…4a). In fact, in gram-positive bacteria, the S-layer lattice primarily consists of multiple bands arranged helically on the cylindrical part of the cell (10,26). Because the Sap protein is synthesized during the exponential phase (20), the ribbon-like appearance could also be due to the fact that this S-layer builds up during the exponential phase, during which the length of the bacterium increases.…”

Section: Discussionmentioning

confidence: 99%

Structural Analysis and Evidence for Dynamic Emergence of Bacillus anthracis S-Layer Networks

Couture‐Tosi¹,

Delacroix

Mignot

et al. 2002

J Bacteriol

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Surface layers (S-layers), which form the outermost layers of many Bacteria and Archaea, consist of protein molecules arranged in two-dimensional crystalline arrays. Bacillus anthracis, a gram-positive, spore-forming bacterium, responsible for anthrax, synthesizes two abundant surface proteins: Sap and EA1. Regulatory studies showed that EA1 and Sap appear sequentially at the surface of the parental strain. Sap and EA1 can form arrays. The structural parameters of S-layers from mutant strains (EA1 ؊ and Sap ؊ ) were determined by computer image processing of electron micrographs of negatively stained regular S-layer fragments or deflated whole bacteria. Sap and EA1 projection maps were calculated on a p1 symmetry basis. The unit cell parameters of EA1 were a ‫؍‬ 69 Å, b ‫؍‬ 83 Å, and ␥ ‫؍‬ 106°, while those of Sap were a ‫؍‬ 184 Å, b ‫؍‬ 81 Å, and ␥ ‫؍‬ 84°. Freeze-etching experiments and the analysis of the peripheral regions of the cell suggested that the two S-layers have different settings. We characterized the settings of each network at different growth phases. Our data indicated that the scattered emergence of EA1 destabilizes the Sap S-layer.

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Section: Discussionmentioning

confidence: 99%

Structural Analysis and Evidence for Dynamic Emergence of Bacillus anthracis S-Layer Networks

Couture‐Tosi¹,

Delacroix

Mignot

et al. 2002

J Bacteriol

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“…New S-layer protein must self-assemble into select sites (11,14,41) to accommodate wall expansion in such a way as to preserve the correct spatial contact with the underlying wall. Somehow, wall turnover, wall expansion, and S-layer growth must be integrated together to preserve contact sites; it seems reasonable to suggest that the same select sites would be used for the growth of both structural layers.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a dynamic S layer on Bacillus thuringiensis

Luckevich

Beveridge

1989

J Bacteriol

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The surfaces of three BaciUus thuringiensis strains possess an S layer composed of linear arrays of small particles arranged with p2 symmetry and with a = 8.5 nm, b = 7.2 nm, and y = 73°. Platinum shadows of whole cells and S-layer fragments revealed the outer surface of the array to be smooth and the inner surface to be corrugated. Treatment with 2 M guanidine hydrochloride at pH 2.5 to 4 best removed the S layer for chemical characterization; it was a relatively hydrophilic 91.4-kilodalton protein with a pl of 5, no detectable carbohydrate, cysteine, methionine or tryptophan, and 21.2% nonpolar residues. No N-terminal homology with other S-layer proteins was evident. Antibody labeling experiments confirmed that the amount of S layer was proportional to the growth phase in broth cultures. Late-exponential-and stationary-growth-phase cells typically sloughed off fragments of S layer, and this may be the result of wall turnover. Indigenous autolytic activity in isolated walls rapidly digested the wall fabric, liberating soluble S-layer protein. At the same time, proteases frequently reduced the molecular weight of the 91.4-kilodalton protein, but these polypeptides could still be identified as S-layer components by immunoblotting. As cultures were serially subcultured, the frequency of appearance of the S layer diminished, and it was eventually lost. The dynamic nature of this S layer makes it atypical of most previously identified S layers and made it unusually difficult to characterize.Our increasing awareness of S layers, or surface arrays, as the outermost component of many archaebacteria and eubacteria (1,13,21,(35)(36)(37)40) exemplifies their possible importance for the vitality and integrity of free-living bacterial cells. Certainly, their importance as the single wall layer of Methanococcus, Thermoproteus, Sulfolobus, and Halobacterium spp. (1, 3, 38) cannot be denied, since this is all that stands between these bacteria and their external milieu.

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“…Pushed or partially masked by growing and fusing crystallites of the newly synthesized S-layer protein, the long teichoic acid molecules protruding from the peptidoglycan layer could form areas of high negative charge in the closing S-layer. For two Bacillus species (Gruber & Sleytr, 1988;Howard et al, 1982), the incorporation of newly synthesized S-layer protein into a coherent S-layer lattice has been shown to be non-random. However, the S-layer reformation described here was a repair process of cells stripped of the S-layer and, thus, probably reflected conditions quite different from the incorporation of S-layer domains in an existing lattice during cell growth.…”

Section: ;mentioning

confidence: 99%

S-layer of Lactobacillus helveticus ATCC 12046: isolation, chemical characterization and re-formation after extraction with lithium chloride

Lortal¹,

Heijenoort²,

Gruber³

et al. 1992

Journal of General Microbiology

140

135

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Localized insertion of new S-layer during growth of Bacillus stearothermophilus strains

Cited by 23 publications

References 27 publications

Structural Analysis and Evidence for Dynamic Emergence of Bacillus anthracis S-Layer Networks

Structural Analysis and Evidence for Dynamic Emergence of Bacillus anthracis S-Layer Networks

Characterization of a dynamic S layer on Bacillus thuringiensis

S-layer of Lactobacillus helveticus ATCC 12046: isolation, chemical characterization and re-formation after extraction with lithium chloride

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