The spoIIN279(ts) mutation affects the FtsA protein of Bacillus subtilis

Karmazyn-Campelli, Céline; Fluss, Larry; Leighton, Terrance; Stragier, Patrick

doi:10.1016/0300-9084(92)90141-z

Cited by 47 publications

(44 citation statements)

References 38 publications

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“…To generate the complementation construct pJS19, a PCR fragment containing the txpA-yqbM intergenic region sequence was amplified with oligonucleotide primers oJS300 and oJS301, ligated to the EcoRI-HindIII sites of pDG364 to place with fragment between the arms of amyE (19), and transformed into JS178 to create JS182. A second complementation construct (pJS63) was generated as described above by PCR amplification of a fragment containing ratA, txpA, and 90 bp of the upstream gene yqbN with oligonucleotide primers oJS425 and oJS426 and ligation to the EcoRI-BamHI sites of pDG364.…”

Section: Methodsmentioning

confidence: 99%

Small Untranslated RNA Antitoxin inBacillus subtilis

Perkins²,

2005

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Toxin-antitoxin (TA) modules are pairs of genes in which one member encodes a toxin that is neutralized or whose synthesis is prevented by the action of the product of the second gene, an antitoxin, which is either protein or RNA. We now report the identification of a TA module in the chromosome of Bacillus subtilis in which the antitoxin is an antisense RNA. The antitoxin, which is called RatA (for RNA antitoxin A), is a small (222 nucleotides), untranslated RNA that blocks the accumulation of the mRNA for a toxic peptide TxpA (for toxic peptide A; formerly YqdB). The txpA and ratA genes are in convergent orientation and overlap by ca. 75 nucleotides, such that the 3 region of ratA is complementary to the 3 region of txpA. Deletion of ratA led to increased levels of txpA mRNA and lysis of the cells. Overexpression of txpA also caused cell lysis and death, a phenotype that was prevented by simultaneous overexpression of ratA. We propose that the ratA transcript is an antisense RNA that anneals to the 3 end of the txpA mRNA, thereby triggering its degradation.

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Section: Methodsmentioning

confidence: 99%

Small Untranslated RNA Antitoxin inBacillus subtilis

Perkins²,

2005

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“…Construction of mother cell-specific and forespore-specific vectors. The SpeI and SacII sites were removed from plasmid pDG364 (17) by digestion with these enzymes, treatment with the Klenow fragment of DNA polymerase and nucleoside triphosphates, and then ligation to create pDPV92. This plasmid contains a chloramphenicol resistance gene and amyE sequences for recombination into the B. subtilis chromosome at the nonessential amyE locus.…”

Section: Methodsmentioning

confidence: 99%

Production of Muramic δ-Lactam in Bacillus subtilis Spore Peptidoglycan

et al. 2004

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Bacterial spore heat resistance is primarily dependent upon dehydration of the spore cytoplasm, a state that is maintained by the spore peptidoglycan wall, the spore cortex. A peptidoglycan structural modification found uniquely in spores is the formation of muramic ␦-lactam. Production of muramic ␦-lactam in Bacillus subtilis requires removal of a peptide side chain from the N-acetylmuramic acid residue by a cwlD-encoded muramoyl-L-Alanine amidase. Expression of cwlD takes place in both the mother cell and forespore compartments of sporulating cells, though expression is expected to be required only in the mother cell, from which cortex synthesis derives. Expression of cwlD in the forespore is in a bicistronic message with the upstream gene ybaK. We show that ybaK plays no apparent role in spore peptidoglycan synthesis and that expression of cwlD in the forespore plays no significant role in spore peptidoglycan formation. Peptide cleavage by CwlD is apparently followed by deacetylation of muramic acid and lactam ring formation. The product of pdaA (yfjS), which encodes a putative deacetylase, has recently been shown to also be required for muramic ␦-lactam formation. Expression of CwlD in Escherichia coli results in muramoyl L-Alanine amidase activity but no muramic ␦-lactam formation. Expression of PdaA alone in E. coli had no effect on E. coli peptidoglycan structure, whereas expression of CwlD and PdaA together resulted in the formation of muramic ␦-lactam. CwlD and PdaA are necessary and sufficient for muramic ␦-lactam production, and no other B. subtilis gene product is required. PdaA probably carries out both deacetylation and lactam ring formation and requires the product of CwlD activity as a substrate.Bacterial endospores can maintain a dormant, highly resistant state for long periods and then, under favorable conditions, rapidly germinate to produce vegetative cells. Spore dormancy and heat resistance are dependent on the relative dehydration of the spore core (6, 24, 28). Spore core dehydration requires the integrity of a thick spore peptidoglycan wall, and peptidoglycan hydrolysis is required for rehydration and resumption of spore core metabolism during spore germination (27,31).The spore peptidoglycan is composed of two contiguous layers that are synthesized between the two membranes surrounding the developing forespore. The inner layer, the germ cell wall, makes up only 10 to 15% of the total spore peptidoglycan (23) and is apparently synthesized by proteins expressed on the surface of the inner forespore membrane (22, 35). The germ cell wall has a structure resembling the peptidoglycan of the vegetative cell wall (23), is maintained during spore germination to serve as the initial wall of the outgrowing spore (5), and appears to function as a template for proper synthesis of the outer spore peptidoglycan layers, the cortex (22). The cortex makes up Ͼ80% of the spore peptidoglycan (23), is synthesized by proteins present on the surface of the outer forespore membrane (10, 35), is rapidly degraded...

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“…The ftsAZ operon was amplified with an EcoRI site at the 5Ј end and a BamHI site at the 3Ј end. Next, it was ligated into the amyE integration vector pDG364 (24), which had been digested with BamHI and EcoRI, thereby inserting the operon into pDG364 in the orientation opposite that of amyE and with the chloramphenicol resistance cassette upstream of the ftsAZ operon. The resulting construct was linearized and transformed directly into B. subtilis RL1075 as above, again bypassing E. coli.…”

Section: Strain Jkb106 ([Py79] Ftsaz::kan Amye::ftsaz[cat]mentioning

confidence: 99%

“…Prior to this study, a mutation of ftsA (originally referred to as spoIIN279 and causing the substitution of an asparagine at Ser9) was known that impairs sporulation (24,31). In contrast to FtsA D265G , which is impaired in gene transcription at or after the stage of polar division, the FtsA S9N mutant is defective in Spo0A-controlled transcription at the onset of sporulation.…”

Section: Isolation Of Ftsa and Ftsz Mutants Impaired In Sporulationmentioning

confidence: 99%

FtsA Mutants ofBacillus subtilisImpaired in Sporulation

2002

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Spore formation in Bacillus subtilis involves a switch in the site of cell division from the midcell to a polar position. Both medial division and polar division are mediated in part by the actin-like, cytokinetic protein FtsA. We report the isolation of an FtsA mutant (FtsA D265G ) that is defective in sporulation but is apparently unimpaired in vegetative growth. Sporulating cells of the mutant reach the stage of asymmetric division but are partially blocked in the subsequent morphological process of engulfment. As judged by fluorescence microscopy and electron microscopy, the FtsA D265G mutant produces normal-looking medial septa but immature (abnormally thin) polar septa. The mutant was unimpaired in transcription under the control of Spo0A, the master regulator for entry into sporulation, but was defective in transcription under the control of F , a regulatory protein whose activation is known to depend on polar division. An amino acid substitution at a residue (Y264) adjacent to D265 also caused a defect in sporulation. D265 and Y264 are conserved among endospore-forming bacteria, raising the possibility that these residues are involved in a sporulation-specific protein interaction that facilitates maturation of the sporulation septum and the activation of F .Most bacteria divide by binary fission, forming a septum at the midpoint of the long axis of the cell to produce identicalsized progeny. The gram-positive soil bacterium Bacillus subtilis exhibits an additional mode of division when it enters the pathway to sporulate. Under such conditions, it undergoes a process of asymmetric division in which a septum is formed at a polar rather than a medial position. The polar septum divides the cell asymmetrically into a forespore (the smaller cell) and a mother cell. The forespore and the mother cell each receive a chromosome but exhibit dissimilar programs of gene expression (reviewed in reference 37). Gene expression in the forespore is governed by the transcription factor F , whose activation depends on the formation of the polar septum, whereas gene expression in the mother cell depends on the transcription factor E . Following asymmetric division, the forespore is engulfed by the mother cell in a phagocyte-like process that results in the forespore becoming pinched off as a free protoplast within the mother cell.The switch from medial division during growth to asymmetric division during sporulation involves a change in the subcellular localization of the cytokinetic protein FtsZ. This tubulinlike GTPase assembles into a ring-like structure, called the Z-ring, at the future site of septation (7, 49). During vegetative growth, the Z-ring assembles at the middle of the cell, fixing the position of the division site. The Z-ring is disassembled during cytokinesis and is largely absent from the completed septum. During sporulation, the Z-ring switches to a bipolar pattern of localization, forming a ring near each pole of the cell (29). Recent evidence indicates that the switch from medial to bipolar Z-rings occur...

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The spoIIN279(ts) mutation affects the FtsA protein of Bacillus subtilis

Cited by 47 publications

References 38 publications

Small Untranslated RNA Antitoxin inBacillus subtilis

Small Untranslated RNA Antitoxin inBacillus subtilis

Production of Muramic δ-Lactam in Bacillus subtilis Spore Peptidoglycan

FtsA Mutants ofBacillus subtilisImpaired in Sporulation

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