Characterization of ywhE, which encodes a putative high-molecular-weight class A penicillin-binding protein in Bacillus subtilis

Pedersen, Lotte B.; Ragkousi, Katerina; Cammett, Tobin J.; Melly, Elizabeth; Sekowska, Agnieszka; Schopick, E.; Murray, Thomas; Setlow, Peter

doi:10.1016/s0378-1119(00)00084-6

Cited by 26 publications

(41 citation statements)

References 26 publications

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“…From the septum, these proteins followed the mother cell membrane during engulfment and ended up in the outer prespore membrane (169). Localization to the prespore is consistent with the role of PBP2c and -2d in sporulation, although it has to be noted that normally these proteins are expressed predominantly in the prespore and not in the mother cell (116,143,156). When GFP-PBP2c was expressed in the mother cell from a promoter that is activated only after the sporulation septum has been closed, the same targeting to the prespore septum and engulfing membrane was observed.…”

Section: Localization Of Pbpssupporting

confidence: 61%

Bacterial Cell Wall Synthesis: New Insights from Localization Studies

Scheffers

Pinho

2005

Microbiol Mol Biol Rev

539

489

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SUMMARY In order to maintain shape and withstand intracellular pressure, most bacteria are surrounded by a cell wall that consists mainly of the cross-linked polymer peptidoglycan (PG). The importance of PG for the maintenance of bacterial cell shape is underscored by the fact that, for various bacteria, several mutations affecting PG synthesis are associated with cell shape defects. In recent years, the application of fluorescence microscopy to the field of PG synthesis has led to an enormous increase in data on the relationship between cell wall synthesis and bacterial cell shape. First, a novel staining method enabled the visualization of PG precursor incorporation in live cells. Second, penicillin-binding proteins (PBPs), which mediate the final stages of PG synthesis, have been localized in various model organisms by means of immunofluorescence microscopy or green fluorescent protein fusions. In this review, we integrate the knowledge on the last stages of PG synthesis obtained in previous studies with the new data available on localization of PG synthesis and PBPs, in both rod-shaped and coccoid cells. We discuss a model in which, at least for a subset of PBPs, the presence of substrate is a major factor in determining PBP localization.

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Section: Localization Of Pbpssupporting

confidence: 61%

Bacterial Cell Wall Synthesis: New Insights from Localization Studies

Scheffers

Pinho

2005

Microbiol Mol Biol Rev

539

489

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“…PBP2d expression is specific to the prespore (Pedersen et al, 2000), but when expressed as a GFP fusion protein during vegetative growth, GFP-PBP2d localized in a dispersed fashion along the membrane, similar to various other PBPs described earlier . To follow GFP-PBP2d during sporulation, it was necessary to keep xylose present in the sporulation medium.…”

Section: Discussionsupporting

confidence: 73%

Dynamic localization of penicillin-binding proteins during spore development in Bacillus subtilis

Scheffers

2005

Microbiology

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During Bacillus subtilis spore formation, many membrane proteins that function in spore development localize to the prespore septum and, subsequently, to the outer prespore membrane. Recently, it was shown that the cell-division-specific penicillin-binding proteins (PBPs) 1 and 2b localize to the asymmetric prespore septum. Here, the author studied the localization of other PBPs, fused to green fluorescent protein (GFP), during spore formation. Fusions to PBPs 4, 2c, 2d, 2a, 3, H, 4b, 5, 4a, 4* and X were expressed during vegetative growth, and their localization was monitored during sporulation. Of these PBPs, 2c, 2d, 4b and 4* have been implicated as having a function in sporulation. It was found that PBP2c, 2d and X changed their localization, while the other PBPs tested were not affected. The putative endopeptidase PbpX appears to spiral out in a pattern that resembles FtsZ redistribution during sporulation, but a pbpX knockout strain had no distinguishable phenotype. PBP2c and 2d localize to the prespore septum and follow the membrane during engulfment, and so are redistributed to the prespore membrane. A similar pattern was observed when GFP-PBP2c was expressed in the mother cell from a sporulation-specific promoter. This work shows that various PBPs known to function during sporulation are redistributed from the cytoplasmic membrane to the prespore.

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“…PBP 2d is not highly expressed during vegetative growth (28) and is therefore not visualized in the PBP profile of vegetative cultures. We found that the band intensities of PBPs 2c, 3, and 4 did not change relative to each other in any strain; however, the band intensities of two class B PBPs, 2a and 2b, did change in relation to each other and to PBP 3.…”

Section: Resultsmentioning

confidence: 97%

Peptidoglycan Synthesis in the Absence of Class A Penicillin-Binding Proteins in Bacillus subtilis

2003

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Penicillin-binding proteins (PBPs) catalyze the final, essential reactions of peptidoglycan synthesis. Three classes of PBPs catalyze either trans-, endo-, or carboxypeptidase activities on the peptidoglycan peptide side chains. Only the class A high-molecular-weight PBPs have clearly demonstrated glycosyltransferase activities that polymerize the glycan strands, and in some species these proteins have been shown to be essential. The Bacillus subtilis genome sequence contains four genes encoding class A PBPs and no other genes with similarity to their glycosyltransferase domain. A strain lacking all four class A PBPs has been constructed and produces a peptidoglycan wall with only small structural differences from that of the wild type. The growth rate of the quadruple mutant is much lower than those of strains lacking only three of the class A PBPs, and increases in cell length and frequencies of wall abnormalities were noticeable. The viability and wall production of the quadruple-mutant strain indicate that a novel enzyme can perform the glycosyltransferase activity required for peptidoglycan synthesis. This activity was demonstrated in vitro and shown to be sensitive to the glycosyltransferase inhibitor moenomycin. In contrast, the quadruple-mutant strain was resistant to moenomycin in vivo. Exposure of the wild-type strain to moenomycin resulted in production of a phenotype similar to that of the quadruple mutant.The structural element of the bacterial cell wall is peptidoglycan (PG), and the importance of this structure is apparent in the number of antimicrobial agents targeting its components and the enzymatic reactions leading to its synthesis. The glycosyltransferase (GT) activity carrying out one of the final enzymatic reactions in PG synthesis is an attractive target, because the enzymes performing that activity are highly conserved and believed to be essential (9, 10). PG is composed of glycan strands cross-linked by peptide side chains. The glycan strands are synthesized by a GT that adds lipid-linked disaccharide pentapeptide subunits to nascent glycan strands. The pentapeptides are then utilized by a transpeptidase to crosslink adjacent glycan strands (reviewed in reference 4). Most of the proteins involved in these final enzymatic reactions are penicillin-binding proteins (PBPs), which are divided into three classes based on the presence of conserved functional domains (9, 10). Only the class A PBPs have an N-terminal domain that contains the conserved amino acid sequences found in all GTs clearly demonstrated to polymerize PG. Class B PBPs have a different N-terminal domain; and although some researchers have associated GT activity with class B PBPs (15), others have been unable to reproduce those results (2). Both classes have C-terminal penicillin-binding domains containing the transpeptidase activity that cross-links peptide side chains. Some species also have monofunctional glycosyltransferases (MGTs) that contain the conserved amino acid sequences found in the class A PBP GT domain but whic...

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Characterization of ywhE, which encodes a putative high-molecular-weight class A penicillin-binding protein in Bacillus subtilis

Cited by 26 publications

References 26 publications

Bacterial Cell Wall Synthesis: New Insights from Localization Studies

Bacterial Cell Wall Synthesis: New Insights from Localization Studies

Dynamic localization of penicillin-binding proteins during spore development in Bacillus subtilis

Peptidoglycan Synthesis in the Absence of Class A Penicillin-Binding Proteins in Bacillus subtilis

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