Transcriptional regulation and structure of the Bacillus subtilis sporulation locus spoIIIC

Errington, Jeff; Rong, Sing; Rosenkrantz, M S; Sonenshein, Abraham L.

doi:10.1128/jb.170.3.1162-1167.1988

Cited by 28 publications

(13 citation statements)

References 40 publications

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“…The complete sequence of each protein was derived from the same sources as those in the legend to Fig. 10, except for Sin (14) and SpollIC (11). data banks, no closely related homologs of SenS were found in other organisms.…”

Section: Methodsmentioning

confidence: 99%

Complex character of senS, a novel gene regulating expression of extracellular-protein genes of Bacillus subtilis

Wang

Doi

1990

J Bacteriol

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The senS gene of Bacillus subtilis, which in high copy number stimulates the expression of several extracellular-protein genes, has been cloned, genetically mapped, and sequenced. The gene codes for a highly charged basic protein containing 65 amino acid residues. The gene is characterized by the presence of a transcription terminator (attenuator) located between the promoter and open reading frame, a strong ribosome-binding site, and a strong transcription terminator at the 3' end of this monocistronic gene. The amino acid sequence of SenS showed partial homology with the N-terminal core binding domain region of bacterial RNA polymerase sigma factors and a helix-turn-helix motif found in DNA-binding proteins. The gene can be deleted without any effect on growth or sporulation.The efficient expression of extracellular-protein genes involves a complex set of transcriptional regulatory factors. These include the products of genes such as sacU (18,24), sacQ (1,24,50), sacV (26), prtR (29,43,52), hpr (19,33) subtilis strains used in this study were the same as those described previously (49). JM101 was used for production of lacZ fusion protein in Escherichia coli. Plasmid pMC1871 was from Pharmacia. pRF373 was a kind gift from Reinhold Bruchner (4). The 2 x SG medium (23) was used as the standard sporulation medium for most of the expression studies.Genetic mapping by using PBS1 transduction. To genetically map the B. natto senN homologous locus in the B.subtilis chromosome, the EcoRI 1.8-kilobase (kb) fragment containing the senN gene (48, 49) was subcloned into the integration plasmid pCP115 (36) to create pCP115-N1.8. This plasmid was then used to transform B. subtilis DB2 (trpC2).Chloramphenicol-resistant (Cmr) transformants were selected on tryptose blood agar base plates containing 5 ,ug of chloramphenicol per p.l. One of the transformants was picked and named DB39. The integration of pCP115-N1.8 at the homologous senS locus was further confirmed by Southern blot analysis. A PBS1 lysate was then prepared from this strain and used to transduce the BGSC mapping-kit strains (7) to Cmr. Molecular cloning of senS by gene conversion. With the previous knowledge of the strong sequence homology between senN and senS as revealed by Southern blot analyses (49) and the homologous integration described above, we used the gene conversion strategy to clone the B. subtilis senS gene. To do this, the N-and C-terminal parts of the senN coding regions were deleted to form plasmids pWL71 and pWL72 (Fig. 1)

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Section: Methodsmentioning

confidence: 99%

Complex character of senS, a novel gene regulating expression of extracellular-protein genes of Bacillus subtilis

Wang

Doi

1990

J Bacteriol

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“…However, the gene encoding K was unknown. Genetic studies focused on the spoIIIC locus, which appeared to encode a small protein with high similarity to the C-terminal region of bacterial factors but lacking any corresponding N-terminal region (64). Interestingly, a different gene, spoIVCB, was found to encode a protein similar to the N-terminal region of a factor.…”

Section: Developmental Chromosome Rearrangementmentioning

confidence: 99%

Compartmentalization of Gene Expression duringBacillus subtilisSpore Formation

Hilbert

Piggot

2004

Microbiol Mol Biol Rev

320

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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“…Recently, Stragier et al (23) showed that spoIVC cisB and spoIIIC (4) are fused at T3 as a result of DNA rearrangement and that the fused gene encodes the late-sporulation-specific sigma factor K (CrK) (15).…”

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confidence: 99%

The cisA cistron of Bacillus subtilis sporulation gene spoIVC encodes a protein homologous to a site-specific recombinase

1990

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The nucleotide sequence of the sporulation gene spolVC cisA in BacUlus subtilis was determined and found to encode a protein of 500 amino acid residues with a calculated molecular weight of 57,481, which It is well known that DNA rearrangement occurs in some cases during cellular differentiation. The best known developmentally regulated rearrangements are those of immunoglobulin genes in vertebrates (24). Since endospore formation of Bacillus subtilis is a simple form of cellular differentiation, it has been speculated for a long time that DNA rearrangement occurs during sporulation (14). Sporulation of B. subtilis initiates in response to nutrient deprivation. In the late stage of sporulation (stages 3 to 7), the cells are divided into two compartments, mother cell and forespore. Each compartment contains a single intact chromosome. The forespore becomes the mature spore, but the mother cell lyses after maturation.The sporulation gene spoIVC of B. subtilis is located in a 3.6-kilobase (kb) EcoRI fragment cloned in temperate bacteriophage 4105 (6). Farquhar and Yudkin (5) described a series of new spoIVC sporulation mutations and suggested that all the mutations fall into two cistrons, cisA and cisB. We found that spoIVC133 is a mutation in the PvuII B fragment using a transformation experiment (6) and that this mutation is located in the cisA gene (see Fig. 1). Kunkel et al. (16) suggested that spoIVC23 is located in cisB and that cisB is expressed from 3.5 h after the initiation of sporulation (T3.5).Recently, Stragier et al. (23) showed that spoIVC cisB and spoIIIC (4) are fused at T3 as a result of DNA rearrangement and that the fused gene encodes the late-sporulation-specific sigma factor K (CrK) (15).We show here that spoIVC cisA encodes a protein whose N-terminal region is highly homologous to DNA recombinases and that the DNA rearrangement does not occur in spoIVC cisA mutants. These results suggest that the spoIVC cisA protein is a site-specific DNA recombinase which rearranges mother cell DNA during sporulation.

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Transcriptional regulation and structure of the Bacillus subtilis sporulation locus spoIIIC

Cited by 28 publications

References 40 publications

Complex character of senS, a novel gene regulating expression of extracellular-protein genes of Bacillus subtilis

Complex character of senS, a novel gene regulating expression of extracellular-protein genes of Bacillus subtilis

Compartmentalization of Gene Expression duringBacillus subtilisSpore Formation

The cisA cistron of Bacillus subtilis sporulation gene spoIVC encodes a protein homologous to a site-specific recombinase

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