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A ring-shaped conduit connects the mother cell and forespore during sporulation in Bacillus subtilis.

ABSTRACT: During spore formation in Bacillus subtilis a transenvelope complex is assembled across the double membrane that separates the mother cell and forespore. This complex (called the "A-Q complex") is required to maintain forespore development and is composed of proteins with remote homology to components of type II, III, and IV secretion systems found in Gram-negative bacteria. Here, we show that one of these proteins, SpoIIIAG, which has remote homology to ring-forming proteins found in type III secretion systems, assembles into an oligomeric ring in the periplasmic-like space between the two membranes. Three-dimensional reconstruction of images generated by cryo-electron microscopy indicates that the SpoIIIAG ring has a cup-and-saucer architecture with a 6-nm central pore. Structural modeling of SpoIIIAG generated a 24-member ring with dimensions similar to those of the EM-derived saucer. Point mutations in the predicted oligomeric interface disrupted ring formation in vitro and impaired forespore gene expression and efficient spore formation in vivo. Taken together, our data provide strong support for the model in which the A-Q transenvelope complex contains a conduit that connects the mother cell and forespore. We propose that a set of stacked rings spans the intermembrane space, as has been found for type III secretion systems.

SUBMITTER: Rodrigues CD

PROVIDER: S-EPMC5068255 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature

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A ring-shaped conduit connects the mother cell and forespore during sporulation in Bacillus subtilis.

Rodrigues Christopher D A CD Henry Xavier X Neumann Emmanuelle E Kurauskas Vilius V Bellard Laure L Fichou Yann Y Schanda Paul P Schoehn Guy G Rudner David Z DZ Morlot Cecile C

Proceedings of the National Academy of Sciences of the United States of America 20160928 41

During spore formation in Bacillus subtilis a transenvelope complex is assembled across the double membrane that separates the mother cell and forespore. This complex (called the "A-Q complex") is required to maintain forespore development and is composed of proteins with remote homology to components of type II, III, and IV secretion systems found in Gram-negative bacteria. Here, we show that one of these proteins, SpoIIIAG, which has remote homology to ring-forming proteins found in type III s ...[more]

PMID: 27681621

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Project description:In the mother cell of sporulating Bacillus subtilis, a regulatory network functions to control gene expression. Four transcription factors act sequentially in the order sigma(E), SpoIIID, sigma(K), GerE. sigma(E) and sigma(K) direct RNA polymerase to transcribe different regulons. SpoIIID and GerE are DNA-binding proteins that activate or repress transcription of many genes. Several negative regulatory loops add complexity to the network. First, transcriptionally active sigma(K) RNA polymerase inhibits early sporulation gene expression, resulting in reduced accumulation of sigma(E) and SpoIIID late during sporulation. Second, GerE represses sigK transcription, reducing sigma(K) accumulation about twofold. Third, SpoIIID represses cotC, which encodes a spore coat protein, delaying its transcription by sigma(K) RNA polymerase. Partially circumventing the first feedback loop, by engineering cells to maintain the SpoIIID level late during sporulation, causes spore defects. Here, the effects of circumventing the second feedback loop, by mutating the GerE binding sites in the sigK promoter region, are reported. Accumulation of pro-sigma(K) and sigma(K) was increased, but no spore defects were detected. Expression of sigma(K)-dependent reporter fusions was altered, increasing the expression of gerE-lacZ and cotC-lacZ and decreasing the expression of cotD-lacZ. Because these effects on gene expression were opposite those observed when the SpoIIID level was maintained late during sporulation, cells were engineered to both maintain the SpoIIID level and have elevated sigK expression late during sporulation. This restored the expression of sigma(K)-dependent reporters to wild-type levels, and no spore defects were observed. Hence, circumventing the second feedback loop suppressed the effects of perturbing the first feedback loop. By feeding information back into the network, these two loops appear to optimize target gene expression and increase network robustness. Circumventing the third regulatory loop, by engineering cells to express cotC about 2 h earlier than normal, did not cause a detectable spore defect.

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A ring-shaped conduit connects the mother cell and forespore during sporulation in Bacillus subtilis.

Publications

A ring-shaped conduit connects the mother cell and forespore during sporulation in Bacillus subtilis.

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