Integrated mapping and sequencing of a 115 kb DNA fragment from Bacillus subtilis: sequence analysis of a 21 kb segment containing the sigL locus

Fabret, Céline; Quentin, Yves; Chapal, Nicolas; Guiseppi, Annick; Haiech, Jacques; Denizot, François

doi:10.1099/13500872-142-11-3089

Cited by 6 publications

(7 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Within the framework of the European Bacillus subtilis genome sequencing project (Moszer et al ., 1996), the chromosomal region between the hisA and sigL loci (approximately 300° on the genetic map) was assigned to the group of F. Denizot and J. Haiech (Laboratoire de Chimie Bactérienne, Marseilles, France) (Fabret et al ., 1996). The nucleotide sequence of a segment from this region revealed an open reading frame (ORF) beginning with an ATG translation initiation codon and ending with a TAA stop codon.…”

Section: Resultsmentioning

confidence: 99%

ClpP ofBacillus subtilisis required for competence development, motility, degradative enzyme synthesis, growth at high temperature and sporulation

Msadek

Dartois

Kunst

et al. 1998

Molecular Microbiology

205

241

View full text Add to dashboard Cite

SummaryThe nucleotide sequence of the Bacillus subtilis clpP gene was determined. The predicted protein shows very high similarity to members of the ClpP family of proteolytic subunits (68% amino acid sequence identity with that of Escherichia coli ). We show that ClpP plays an essential role in stationary phase adaptive responses. Indeed, a ⌬clpP mutant was constructed and shown to display a pleiotropic phenotype, including a deficiency in both sporulation initiation and competence for DNA uptake. The ⌬clpP mutant has a highly filamentous morphology and appears to be non-motile, as judged by swarm plate assays. Expression of clpP is strongly induced under heat shock conditions, and ClpP is shown to be essential for growth of B. subtilis at high temperature. The role of ClpP in the sporulation and competence regulatory pathways was investigated. ClpP is required for expression of the spoIIA and spoIIG operons, encoding the F and E sporulation-specific sigma factors. ClpP is also necessary for the expression of the comK gene, encoding a positive transcriptional regulator of competence genes. ComK-dependent transcription of sacB, encoding the exocellular degradative enzyme levansucrase, was found to be abolished in the ⌬clpP mutant. MecA has been characterized previously as a negative regulator of comK expression, whose overproduction inhibits both sporulation and competence development. Expression of a mecAЈ-ЈlacZ translational fusion is shown to be increased in the ⌬clpP mutant. We suggest that ClpP is involved in controlling MecA levels in the cell through proteolysis. Increased levels of MecA in the absence of ClpP are at least partly responsible for the observed pleiotropic phenotype of the ⌬clpP mutant.

show abstract

Section: Resultsmentioning

confidence: 99%

ClpP ofBacillus subtilisis required for competence development, motility, degradative enzyme synthesis, growth at high temperature and sporulation

Msadek

Dartois

Kunst

et al. 1998

Molecular Microbiology

205

241

View full text Add to dashboard Cite

show abstract

“…Inositol dehydrogenase activities were determined as described (9) by measuring the formation of NADH from NAD plus inositol at 37°C. (32) and encoding a protein composed of 85 amino acids has been identified. This protein exhibits 45% sequence identity and 67% similarity to HPr (Fig.…”

Section: Methodsmentioning

confidence: 99%

The Bacillus subtilis crh gene encodes a HPr-like protein involved in carbon catabolite repression

Galinier

Haiech

Kilhoffer

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

208

274

View full text Add to dashboard Cite

Carbon catabolite repression (CCR) of several Bacillus subtilis catabolic genes is mediated by ATPdependent phosphorylation of histidine-containing protein (HPr), a phosphocarrier protein of the phosphoenolpyruvate (PEP): sugar phosphotransferase system. In this study, we report the discovery of a new B. subtilis gene encoding a HPr-like protein, Crh (for catabolite repression HPr), composed of 85 amino acids. Crh exhibits 45% sequence identity with HPr, but the active site His-15 of HPr is replaced with a glutamine in Crh. Crh is therefore not phosphorylated by PEP and enzyme I, but is phosphorylated by ATP and the HPr kinase in the presence of fructose-1,6-bisphosphate. We determined Ser-46 as the site of phosphorylation in Crh by carrying out mass spectrometry with peptides obtained by tryptic digestion or CNBr cleavage. In a B. subtilis ptsH1 mutant strain, synthesis of ␤-xylosidase, inositol dehydrogenase, and levanase was only partially relieved from CCR. Additional disruption of the crh gene caused almost complete relief from CCR. In a ptsH1 crh1 mutant, producing HPr and Crh in which Ser-46 is replaced with a nonphosphorylatable alanyl residue, expression of ␤-xylosidase was also completely relieved from glucose repression. These results suggest that CCR of certain catabolic operons requires, in addition to CcpA, ATP-dependent phosphorylation of Crh, and HPr at Ser-46.The bacterial phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS) catalyzes the transport and concomitant phosphorylation of carbohydrates via a protein phosphorylation chain including PEP-dependent phosphorylation of His-15 in histidine-containing protein (HPr) by enzyme I (EI). P-His-HPr phosphorylates the sugar-specific EIIAs. In Gram-positive bacteria, the PTS regulates also induction and carbon catabolite repression (CCR) of numerous catabolic genes (1). The central regulatory protein involved in these various functions is HPr. In Gram-positive bacteria, this small phosphoryl transfer protein can be phosphorylated at a regulatory serine (Ser-46) by ATP and the HPr kinase (2, 3), in addition to phosphorylation at the catalytic His-15 by PEP and EI (4, 5). PEP-dependent and ATP-dependent phosphorylation of HPr interfere with each other-i.e., P-His-HPr is a poor substrate for the HPr kinase and P-Ser-HPr is a poor substrate for EI (6, 7). ATP-dependent phosphorylation of HPr is stimulated by glycolytic intermediates such as fructose-1,6-bisphosphate (FBP) in Enterococcus faecalis (6) and in Streptococcus pyogenes (7). It has been reported that FBP is also implicated in CCR of the Bacillus subtilis gnt and iol operons (8, 9), and a potential role of phosphorylation of HPr at Ser-46 in CCR has therefore been investigated (10). The gnt operon contains the genes gntRKPZ encoding the repressor GntR, gluconate kinase, gluconate permease, and a gluconate-6-Pdehydrogenase (11), whereas the iol operon is composed of 10 genes encoding enzymes presumably implicated in inositol metabolism, including iolG encoding inositol dehydro...

show abstract

“…Some of the functions encoded within these variable regions are also known to have an unusual base composition in other species or are known to vary among strains. These include the exopolysaccharide biosynthesis regions (11,14,17) and the restriction modification systems (38). Some variable regions have a base composition that is similar to the genome average.…”

Section: Fig 2 Conservation and Variability Of Genes In L Plantarumentioning

confidence: 99%

ExploringLactobacillus plantarumGenome Diversity by Using Microarrays

et al. 2005

View full text Add to dashboard Cite

Lactobacillus plantarum is a versatile and flexible species that is encountered in a variety of niches and can utilize a broad range of fermentable carbon sources. To assess if this versatility is linked to a variable gene pool, microarrays containing a subset of small genomic fragments of L. plantarum strain WCFS1 were used to perform stringent genotyping of 20 strains of L. plantarum from various sources. The gene categories with the most genes conserved in all strains were those involved in biosynthesis or degradation of structural compounds like proteins, lipids, and DNA. Conversely, genes involved in sugar transport and catabolism were highly variable between strains. Moreover, besides the obvious regions of variance, like prophages, other regions varied between the strains, including regions encoding plantaricin biosynthesis, nonribosomal peptide biosynthesis, and exopolysaccharide biosynthesis. In many cases, these variable regions colocalized with regions of unusual base composition. Two large regions of flexibility were identified between 2.70 and 2.85 and 3.10 and 3.29 Mb of the WCFS1 chromosome, the latter being close to the origin of replication. The majority of genes encoded in these variable regions are involved in sugar metabolism. This functional overrepresentation and the unusual base composition of these regions led to the hypothesis that they represented lifestyle adaptation regions in L. plantarum. The present study consolidates this hypothesis by showing that there is a high degree of gene content variation among L. plantarum strains in genes located in these regions of the WCFS1 genome. Interestingly, based on our genotyping data L. plantarum strains clustered into two clearly distinguishable groups, which coincided with an earlier proposed subdivision of this species based on conventional methods.

show abstract

Integrated mapping and sequencing of a 115 kb DNA fragment from Bacillus subtilis: sequence analysis of a 21 kb segment containing the sigL locus

Cited by 6 publications

References 32 publications

ClpP ofBacillus subtilisis required for competence development, motility, degradative enzyme synthesis, growth at high temperature and sporulation

ClpP ofBacillus subtilisis required for competence development, motility, degradative enzyme synthesis, growth at high temperature and sporulation

The Bacillus subtilis crh gene encodes a HPr-like protein involved in carbon catabolite repression

ExploringLactobacillus plantarumGenome Diversity by Using Microarrays

Contact Info

Product

Resources

About