The life‐cycle proteins RodA of <i>Escherichia coli</i> and SpoVE of Bacillus subtilis have very similar primary structures

Joris, Bernard; Dive, Georges; Henriques, Adriano O.; Piggot, Patrick J.; Ghuysen, Jean‐Marie

doi:10.1111/j.1365-2958.1990.tb00618.x

Cited by 37 publications

(25 citation statements)

References 44 publications

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“…A similar result was obtained for the B. subtilis rodC operon (6). Recently, it was found that the spoVE gene product is homologous to the newly identified E. coli ftsW gene product, in which mutations induce filamentous cell growth at the nonpermissive temperature, and to the newly sequenced E. coli rodA gene product (7,10). These results suggest that SpoVE or the succeeding gene product MurG (15) plays an essential role not only during sporulation but also during vegetative growth.…”

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confidence: 74%

In vivo expression of the Bacillus subtilis spoVE gene

et al. 1993

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In vivo expression of the Bacillus subtilis spoVE gene was studied by S1 nuclease mapping and spoVE gene fusion analysis. Transcription of spoVE is induced at about the second hour of sporulation from two closely spaced promoters designated P1 and P2. Examination of the precise transcription initiation site by highresolution primer extension mapping indicated that the nucleotide sequences of the -10 and -35 regions of both P1 and P2 were similar to those of promoters recognized by Eo'". Moreover, S1 nuclease mapping and translational spoVE-lacZ fusion studies with various spo mutants suggest that the expression of spoVE P2 requires the spoIlG gene product, &E. The sporulation of a wild-type strain was inhibited severely in the presence of a multicopy plasmid, pKBVE, carrying the spoVE promoter, indicating the possible titration of a transcriptional regulatory element(s).Endospore formation in Bacillus subtilis involves an elaborate program of morphological development (13, 19) which requires the expression of at least 50 genetic loci, some of which are operons containing several sporulation-essential genes. Most of these loci, except for spoO genes, are not expressed before the initiation of sporulation, which is followed by the sequential expression of the sigma factors (25) and the crisscross regulation of cell-type-specific gene expression (12).Previously, one of the stage V sporulation genes, spoVE, has been cloned (18, 28) and the nucleotide sequence has been determined (2, 22). spoVE mutations cause an unusual defect in spore formation in which the forespores are surrounded by well-developed spore coat layers but the cortex is almost entirely absent (19). When the spoVE mRNA synthesis was analyzed by dot blot hybridization with a part of the spoVE structural gene as a probe, it was found that the spoVE mRNA was synthesized from the vegetative-growth phase and that the synthesis was not prevented by early spo mutations (16). On the contrary, when the transcriptional regulation was studied by using the spoVE-lacZ gene fusion, the expression of the hybrid gene could be detected about 40 min after the onset of sporulation (1). It was also observed that mutations in spoOH and spoOK but not in spoIL4 and spoIIG impede the expression of spoVE (1). For the present paper, we investigated the in vivo expression of the spoVE gene by S1 nuclease mapping and spoVE gene fusion experiments. Our results show that the spoVE gene is turned on just before the onset of cortex formation and suggest that the expression of spoVE requires the spoIIG gene products. MATERIALS AND METHODSBacterial strains and plasmids. The bacterial strains and plasmids used in this study are listed in

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Section: Discussionsupporting

confidence: 74%

In vivo expression of the Bacillus subtilis spoVE gene

et al. 1993

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“…Fortunately, a third SEDS family member functions during the nonessential process of sporulation in Bacillus subtilis (18,20). Bacterial endospores can survive extremes of heat and desiccation, largely because of the presence of the spore cortex.…”

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Determinants for the Subcellular Localization and Function of a Nonessential SEDS Protein

et al. 2008

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The Bacillus subtilis SpoVE integral membrane protein is essential for the heat resistance of spores, probably because of its involvement in spore peptidoglycan synthesis. We found that an SpoVE-yellow fluorescent protein (YFP) fusion protein becomes localized to the forespore during the earliest stages of engulfment, and this pattern is maintained throughout sporulation. SpoVE belongs to a well-conserved family of proteins that includes the FtsW and RodA proteins of B. subtilis. These proteins are involved in bacterial shape determination, although their function is not known. FtsW is necessary for the formation of the asymmetric septum in sporulation, and we found that an FtsW-YFP fusion localized to this structure prior to the initiation of engulfment in a nonoverlapping pattern with SpoVE-cyan fluorescent protein. Since FtsW and RodA are essential for normal growth, it has not been possible to identify loss-of-function mutations that would greatly facilitate analysis of their function. We took advantage of the fact that SpoVE is not required for growth to obtain point mutations in SpoVE that block the development of spore heat resistance but that allow normal protein expression and targeting to the forespore. These mutant proteins will be invaluable tools for future experiments aimed at elucidating the function of members of the SEDS ("shape, elongation, division, and sporulation") family of proteins.Bacterial shape is determined by an extracellular structure composed of peptidoglycan (PG), a rigid polymer of repeated subunits of a disaccharide peptide monomer. This giant macromolecule is found on the outside of the cytoplasmic membrane of nearly all eubacteria. A series of essential and highly conserved enzymes convert UDP-GlcNAc to lipid II, the UDPmuramyl pentapeptide (47). To form mature PG, lipid II is translocated across the cytoplasmic membrane via an uncharacterized mechanism and is added to the preexisting cell wall through transpeptidation and transglycosylation reactions mediated by members of the PBP (penicillin binding protein) family of proteins. Little is known about the mechanism of lipid II transport across the cytoplasmic membrane other than that it is probably dependent on a protein(s) since it does not occur spontaneously across lipid bilayers (46).The integral membrane proteins RodA and FtsW are members of what has been termed the SEDS ("shape, elongation, division, and sporulation") family of integral membrane proteins (16, 18) that are present in all cell wall-containing bacteria. Several lines of evidence are consistent with the participation of RodA and FtsW in the translocation of lipid II during cell elongation and cell division, respectively (17, 19), although this hypothesis has not been subjected to a direct test. For instance, depletion of RodA leads to a block in lateral cell growth (16), although RodA is not strictly essential for cell viability. Null mutants exhibit slow growth and small cell diameters and are viable in minimal medium (7). Also, temperature-sensitive Escheri...

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“…For example, inactivation of FtsI delays Z-ring assembly and reduces the total number of rings per unit cell mass by about twofold compared to an unperturbed population of cells (34). Even inactivation of FtsA or ZipA, which localize immediately after FtsZ, has only a modest effect on the number of Z-rings observed per unit mass (2,18,27).FtsW belongs to a large family of polytopic membrane proteins that appear to be present in all bacteria that have a peptidoglycan cell wall (21,22,24). This family has been named SEDS (21) for shape, elongation, division, and sporulation.…”

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The Escherichia coli Cell Division Protein FtsW Is Required To Recruit Its Cognate Transpeptidase, FtsI (PBP3), to the Division Site

2002

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The bacterial cell division protein FtsW has been suggested to perform two functions: stabilize the FtsZ cytokinetic ring, and facilitate septal peptidoglycan synthesis by the transpeptidase FtsI (penicillin-binding protein 3). We show here that depleting Escherichia coli cells of FtsW had little effect on the abundance of FtsZ rings but abrogated recruitment of FtsI to potential division sites. Analysis of FtsW localization confirmed and extended these results; septal localization of FtsW required FtsZ, FtsA, FtsQ, and FtsL but not FtsI. Thus, FtsW is a late recruit to the division site and is essential for subsequent recruitment of its cognate transpeptidase FtsI but not for stabilization of FtsZ rings. We suggest that a primary function of FtsW homologueswhich are found in almost all bacteria and appear to work in conjunction with dedicated transpeptidases involved in division, elongation, or sporulation-is to recruit their cognate transpeptidases to the correct subcellular location.FtsW is an integral membrane protein required for cell division in Escherichia coli (6,23,26), and presumably most other bacteria as well, although its precise biochemical role in the process is not yet known. FtsW is one of nine essential cell division proteins in E. coli, all of which have been shown to localize to the division site (midcell) during septation.Studies of localization in various mutant backgrounds revealed a set of dependency relationships that are generally considered to reflect the order in which the division proteins are recruited to the division site (reviewed in reference 35). The first event in this pathway is assembly of FtsZ into a ring (Z-ring) at the future division site (Fig. 1). FtsA and ZipA each bind directly to FtsZ and localize next. After localization of FtsA, the proteins FtsK, FtsQ, FtsL, FtsI, and FtsN appear at the division site in that order. This recruitment sequence has been postulated to reflect the order of assembly of a multiprotein complex that mediates constriction of the cell envelope at the midcell (28,30,33,35).Although FtsW is known to localize to the division site (43), its position in the pathway has not been established. The most pertinent study reported that depletion of FtsW severely destabilized Z-rings, implying that FtsW is an early recruit to the division site and has an important role in regulation of FtsZ dynamics (6). In contrast, inactivation of other division proteins has only a minor effect on Z-rings. For example, inactivation of FtsI delays Z-ring assembly and reduces the total number of rings per unit cell mass by about twofold compared to an unperturbed population of cells (34). Even inactivation of FtsA or ZipA, which localize immediately after FtsZ, has only a modest effect on the number of Z-rings observed per unit mass (2,18,27).FtsW belongs to a large family of polytopic membrane proteins that appear to be present in all bacteria that have a peptidoglycan cell wall (21,22,24). This family has been named SEDS (21) for shape, elongation, division, and sporulat...

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The life‐cycle proteins RodA of Escherichia coli and SpoVE of Bacillus subtilis have very similar primary structures

Cited by 37 publications

References 44 publications

In vivo expression of the Bacillus subtilis spoVE gene

In vivo expression of the Bacillus subtilis spoVE gene

Determinants for the Subcellular Localization and Function of a Nonessential SEDS Protein

The Escherichia coli Cell Division Protein FtsW Is Required To Recruit Its Cognate Transpeptidase, FtsI (PBP3), to the Division Site

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