Expression, secretion and antigenic variation of bacterial S‐layer proteins

Boot, Hein J.; Pouwels, Peter H.

doi:10.1046/j.1365-2958.1996.711442.x

Cited by 119 publications

(114 citation statements)

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“…Secretion followed by surface localization has been described for surface entities from a range of microbes, both prokaryotes and eukaryotes (3,4,12). Interestingly, some secreted and reattached proteins, like Blastomyces dermatitidis Bad1p and MIC3 of Toxoplasma gondii, contain EGF-like or chitin binding domains (5,12).…”

Section: Discussionmentioning

confidence: 99%

Surface Localization of the Yps3p Protein of Histoplasma capsulatum

Bohse

Woods

2005

Eukaryot Cell

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The YPS3 gene of Histoplasma capsulatum encodes a protein that is both resident in the cell wall and also released into the culture medium. This protein is produced only during the pathogenic yeast phase of infection and is also expressed differently in H. capsulatum strains that differ in virulence. We investigated the cellular localization of Yps3p. We demonstrated that the cell wall fraction of Yps3p was surface localized in restriction fragment length polymorphism class 2 strains. We also established that Yps3p released into the G217B culture supernatant binds to the surface of strains that do not naturally express the protein. This binding was saturable and occurred within 5 min of exposure and occurred similarly with live and heat-killed H. capsulatum. Flow cytometric analysis of H. capsulatum after enzymatic treatments was consistent with Yps3p binding to chitin, a carbohydrate polymer that is a component of fungal cell walls. Polysaccharide binding assays demonstrated that chitin but not cellulose binds to and extracts Yps3p from culture supernatants.

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Section: Discussionmentioning

confidence: 99%

Surface Localization of the Yps3p Protein of Histoplasma capsulatum

Bohse

Woods

2005

Eukaryot Cell

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“…Accordingly, S-layer genes are expressed at extremely high levels, and S-layer proteins constitute 10 to 15 % of the total protein content of the cell (Messner & Sleytr, 1992). The high-level expression of S-layer genes is based not only on strong promoters and efficient transcription but also on highly stable mRNA molecules (Boot & Pouwels, 1996;Fisher et al, 1988;Kahala et al, 1997). However, S-layer proteins are seldom found in culture supernatants, suggesting that their synthesis is tightly regulated.…”

Section: Introductionmentioning

confidence: 99%

The surface (S)-layer gene cspB of Corynebacterium glutamicum is transcriptionally activated by a LuxR-type regulator and located on a 6 kb genomic island absent from the type strain ATCC 13032

et al. 2006

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The surface (S)-layer gene region of the Gram-positive bacterium Corynebacterium glutamicum ATCC 14067 was identified on fosmid clones, sequenced and compared with the genome sequence of C. glutamicum ATCC 13032, whose cell surface is devoid of an ordered S-layer lattice. A 5·97 kb DNA region that is absent from the C. glutamicum ATCC 13032 chromosome was identified. This region includes cspB, the structural gene encoding the S-layer protomer PS2, and six additional coding sequences. PCR experiments demonstrated that the respective DNA region is conserved in different C. glutamicum wild-type strains capable of S-layer formation. The DNA region is flanked by a 7 bp direct repeat, suggesting that illegitimate recombination might be responsible for gene loss in C. glutamicum ATCC 13032. Transfer of the cloned cspB gene restored the PS2− phenotype of C. glutamicum ATCC 13032, as confirmed by visualization of the PS2 proteins by SDS-PAGE and imaging of ordered hexagonal S-layer lattices on living C. glutamicum cells by atomic force microscopy. Furthermore, the promoter of the cspB gene was mapped by 5′ rapid amplification of cDNA ends PCR and the corresponding DNA fragment was used in DNA affinity purification assays. A 30 kDa protein specifically binding to the promoter region of the cspB gene was purified. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and peptide mass fingerprinting of the purified protein led to the identification of the putative transcriptional regulator Cg2831, belonging to the LuxR regulatory protein family. Disruption of the cg2831 gene in C. glutamicum resulted in an almost complete loss of PS2 synthesis. These results suggested that Cg2831 is a transcriptional activator of cspB gene expression in C. glutamicum.

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“…stearothermophilus) (for review, see . These proteins interact with underlying cell-wall polymers via non-covalent bonds (Boot & Pouwels, 1996 ;Ries et al, 1997 ;Mesnage et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

A general strategy for identification of S-layer genes in the Bacillus cereus group: molecular characterization of such a gene in Bacillus thuringiensis subsp. galleriae NRRL 4045 The GenBank accession number for the slpA sequence is AJ249446.

2001

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Despite its possible role in virulence, there has been little molecular characterization of members of the S-layer protein family of the Bacillus cereus group. It is hypothesized that the components of the S-layers are likely to display similar anchoring structures in Bacillus thuringiensis and Bacillus anthracis. Based on inferred sequence similarities, a DNA fragment encoding the cell-wall-anchoring domain of an S-layer component of Bac. thuringiensis subsp. galleriae NRRL 4045 was isolated. The complete gene was identified and sequenced. An ORF of 2463 nt was identified, which was predicted to encode a protein of 821 aa, SlpA. The mature SlpA protein (792 residues) carries three S-layer homology (SLH) motifs towards its amino terminus, each about 50 aa long. These motifs were sufficient to bind Bac. thuringiensis and Bac. anthracis cell walls in vitro by interacting with peptidoglycan-associated polymers, confirming a common wall-anchoring mechanism. The slpA null-allele mutant was constructed and shown to possess no other abundant surface protein. It is proposed that the method described in this paper could be applied to the identification and deletion of any Bac. cereus S-layer gene and is of great value for the molecular and functional characterization of these genes.

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Expression, secretion and antigenic variation of bacterial S‐layer proteins

Cited by 119 publications

References 50 publications

Surface Localization of the Yps3p Protein of Histoplasma capsulatum

Surface Localization of the Yps3p Protein of Histoplasma capsulatum

The surface (S)-layer gene cspB of Corynebacterium glutamicum is transcriptionally activated by a LuxR-type regulator and located on a 6 kb genomic island absent from the type strain ATCC 13032

A general strategy for identification of S-layer genes in the Bacillus cereus group: molecular characterization of such a gene in Bacillus thuringiensis subsp. galleriae NRRL 4045 The GenBank accession number for the slpA sequence is AJ249446.

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