Physical and functional characterization of the Bacillus subtilis spoIIM gene

Smith, K. M.; Bayer, Michaela; Youngman, Philip

doi:10.1128/jb.175.11.3607-3617.1993

Cited by 57 publications

(80 citation statements)

References 62 publications

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“…In the present work, we have investigated the temporal and spatial regulation of the spolIM gene, a gene whose product is required for migration of the asymmetric septum and complete engulfment of the forespore compartment (69) We find clear evidence that spoIIM is a member of the &r regulon, but we find conflicting indications concerning the compartmentalization of its expression.…”

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confidence: 80%

“…product suggests that transcription of the spoIIM gene initiates at a T residue 35 bp upstream from an ATG codon believed to be the start of the spoIIM coding sequence (69). Centered 10 and 32 bases upstream from this presumptive transcriptional start site are sequences that conform almost perfectly to the consensus for promoters recognized by cr-associated RNA polymerase (Fig.…”

mentioning

confidence: 99%

“…Selection for antibiotic resistance in B. subtilis with chloramphenicol (CM), erythromycin-lincomycin, macrolides-lincosamides-streptogramin B (MLS), phleomycin (PM), and kanamycin was as previously described (23,(85)(86)(87). Selection for antibiotic resistance in E. coli with ampicillin and tetracycline was as previously described (69).…”

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Evidence that the spoIIM gene of Bacillus subtilis is transcribed by RNA polymerase associated with sigma E

Smith

Youngman

1993

J Bacteriol

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We have investigated the temporal and spatial regulation ofspoHIM, a gene ofBacillus subtilis whose product is required for complete septum migration and engulfment of the forespore compartment during sporulation. The spoHIM promoter was found to become active about 2 h after the initiation of sporulation. The effects of mutations on the expression of a spoIIM-lacZ fusion were most consistent with its utilization by &-Fassociated RNA polymerase (Eci). A unique 5' end of the in vivo spoHIM transcript was detected by primer extension analysis and was determined to initiate at the appropriate distance from a sequence conforming very closely to the consensus for genes transcribed by E&F. A partially purified preparation of E&X produced a transcript in vitro that initiated at the same nucleotide as the primer extension product generated from in vivo RNA. Ectopic induction of &E synthesis during growth resulted in the immediate and strong expression of a spoIIM-lacZ fusion, but an identical fusion was completely unresponsive to induced synthesis of either oF or rG under similar conditions. The results of plasmid integration-excision experiments in which the spolIM gene was reversibly disrupted by a temperature-sensitive integrational vector suggested that spolIM expression is required in the forespore compartment, but direct examination of subcellular fractions enriched for mother cell or forespore material indicated that spolIM expression cannot be confined to the forespore. We conclude that spoHIM is a member of the &E regulon and that it may be transcribed exclusively by Ecr-. We discuss the implications of this conclusion for models in which activation of o1r in the mother cell is proposed to be a part of the mechanism responsible for initiating separate programs of gene activity in the two sporangium compartments.Endospore formation in Bacillus subtilis involves a series of temporally and spatially ordered changes in cell morphology and gene expression (42,54). At a critical stage in this process, an asymmetric septation event creates two cell compartments, the mother cell and the forespore, each containing a transcriptionally active chromosome. At some point very soon after asymmetric septation, the mother cell and the forespore compartments begin to execute quite distinct programs of gene expression. The mechanisms responsible for establishing these compartment-specific programs are not fully understood, but they apparently involve, in part, the compartment-specific activation of RNA polymerase sigma factors (16,40,41,76). This is most obvious at the later stages of sporulation, in which a polymerase species associated with g is active exclusively in the mother cell compartment (11,36,43) and a species associated with cyG is active exclusively in the forespore (31, 78). At earlier stages, the situation is less certain and the basis for compartmentspecific gene activity is more speculative. Soon after asymmetric septation, a polymerase species associated with &rF becomes active in the forespore (45, 67). The o-F...

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Evidence that the spoIIM gene of Bacillus subtilis is transcribed by RNA polymerase associated with sigma E

Smith

Youngman

1993

J Bacteriol

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“…Expression of both prespore-specific and mother cell-specific loci is required for engulfment. spoIID (37,228), spoIIM (271), and spoIIP (85) mutants display autolysis and membrane flexibility at the center of the spore septum but not its periphery. spoIIQ mutants display a block just prior to completion of engulfment (178).…”

Section: Initiation Of Engulfmentmentioning

confidence: 99%

“…Members of the E regulon that are important for sporulation include spoIID, spoIIM, and spoIIP, which are required for engulfment and to prevent a second asymmetric division from occurring in the mother cell (1,37,57,85,228,232,246,271,272); spoIVA, cotE, and spoVID, which encode scaffold proteins for spore coat assembly (18,245,273,317,318); the spoIIIA operon, which is required for activation of the lateprespore specific sigma factor G (125, 146); sigK, the composite gene for the late mother cell-specific transcription factor K (155,280); and spoIVCA, the gene for the recombinase that generates sigK via a chromosomal rearrangement (155,235,256). In addition, E activates transcription of spoIIID, which encodes a regulator of some E -dependent genes (102).…”

Section: E Regulonmentioning

confidence: 99%

Compartmentalization of Gene Expression duringBacillus subtilisSpore Formation

Hilbert

Piggot

2004

Microbiol Mol Biol Rev

318

432

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Gene expression in members of the family Bacillaceae becomes compartmentalized after the distinctive, asymmetrically located sporulation division. It involves complete compartmentalization of the activities of sporulation-specific sigma factors, σF in the prespore and then σE in the mother cell, and then later, following engulfment, σG in the prespore and then σK in the mother cell. The coupling of the activation of σF to septation and σG to engulfment is clear; the mechanisms are not. The σ factors provide the bare framework of compartment-specific gene expression. Within each σ regulon are several temporal classes of genes, and for key regulators, timing is critical. There are also complex intercompartmental regulatory signals. The determinants for σF regulation are assembled before septation, but activation follows septation. Reversal of the anti-σF activity of SpoIIAB is critical. Only the origin-proximal 30% of a chromosome is present in the prespore when first formed; it takes ≈15 min for the rest to be transferred. This transient genetic asymmetry is important for prespore-specific σF activation. Activation of σE requires σF activity and occurs by cleavage of a prosequence. It must occur rapidly to prevent the formation of a second septum. σG is formed only in the prespore. SpoIIAB can block σG activity, but SpoIIAB control does not explain why σG is activated only after engulfment. There is mother cell-specific excision of an insertion element in sigK and σE-directed transcription of sigK, which encodes pro-σK. Activation requires removal of the prosequence following a σG-directed signal from the prespore

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The engulfasome in C. difficile: Variations on protein machineries

Kelly

Salgado

2019

Anaerobe

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Clostridioides difficile infection (CDI) continues to be a substantial healthcare burden, and the changing disease profile raises new challenges in CDI management, both in clinical settings and in the community. CDI is transmitted by spores, which are formed by a subset of the cell population where an asymmetric septum is formed. A full copy of the chromosome is transported into the smaller compartment which is then engulfed by the mother cell. After engulfment, multiple metabolic and morphological changes occur, eventually resulting in the release of the mature spore. Whilst studies in the model organism Bacillus subtilis have demonstrated the importance of the DMP and Q:AH machineries in engulfment, it is becoming clear that there are fundamental differences in the way the two organisms organise these machineries. As spores are the infectious agent in CDI, it is crucial to understand how these dormant cells are formed, and how sporulation can be prevented or disrupted with the view of reducing CDI. Here, we review the current literature on the DMP and Q:AH machineries in C. difficile, and how they compare and contrast to those of B. subtilis.

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Physical and functional characterization of the Bacillus subtilis spoIIM gene

Cited by 57 publications

References 62 publications

Evidence that the spoIIM gene of Bacillus subtilis is transcribed by RNA polymerase associated with sigma E

Evidence that the spoIIM gene of Bacillus subtilis is transcribed by RNA polymerase associated with sigma E

Compartmentalization of Gene Expression duringBacillus subtilisSpore Formation

The engulfasome in C. difficile: Variations on protein machineries

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