Transcriptional regulation of the Bacillus subtilis menp1 promoter

Xuan, Qin; Taber, Harry

doi:10.1128/jb.178.3.705-713.1996

Cited by 10 publications

(9 citation statements)

References 32 publications

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“…There is a similar situation in the menD mutant because interruption of menadione biosynthesis is accompanied by a coordinate decrease in heme biosynthesis in gram-positive bacteria (7,23). Because the menD mutant is more metabolically restricted, it may be less able to maintain its ⌬⌿, making it more susceptible to these compounds.…”

Section: Discussionmentioning

confidence: 99%

Phenotype Microarray Profiling of Staphylococcus aureus menD and hemB Mutants with the Small-Colony-Variant Phenotype

et al. 2006

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Standard biochemical tests have revealed that hemin and menadione auxotrophic Staphylococcus aureus smallcolony variants (SCVs) exhibit multiple phenotypic changes. To provide a more complete analysis of the SCV phenotype, two genetically defined mutants with a stable SCV phenotype were comprehensively tested. These mutants, generated via mutations in menD or hemB that yielded menadione and hemin auxotrophs, were subjected to phenotype microarray (PM) analysis of over 1,500 phenotypes (including utilization of different carbon, nitrogen, phosphate, and sulfur sources; growth stimulation or inhibition by amino acids and other nutrients, osmolytes, and metabolic inhibitors; and susceptibility to antibiotics). Compared to parent strain COL, the hemB mutant was defective in utilization of a variety of carbon sources, including Krebs cycle intermediates and compounds that ultimately generate ATP via electron transport. The phenotype of the menD mutant was similar to that of the hemB mutant, but the defects in carbon metabolism were more pronounced than those seen with the hemB mutant. In both mutant strains, hexose phosphates and other carbohydrates that provide ATP in the absence of electron transport stimulated growth. Other phenotypes of SCV mutants, such as hypersensitivity to sodium selenite, sodium tellurite, and sodium nitrite, were also uncovered by the PM analysis. Key results of the PM analysis were confirmed in independent growth studies and by using Etest strips for susceptibility testing. PM technology is a new and efficient technology for assessing cellular phenotypes in S. aureus.

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Section: Discussionmentioning

confidence: 99%

Phenotype Microarray Profiling of Staphylococcus aureus menD and hemB Mutants with the Small-Colony-Variant Phenotype

et al. 2006

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“…The menF promoter, menp1, appears to be responsive to the carbon source and the growth phase (28). Transcription of dhbC is controlled by a promoter upstream of dhbA.…”

Section: Vol 178 1996 Isochorismate Synthase Genes Of Bacillus Subtmentioning

confidence: 99%

Duplicate isochorismate synthase genes of Bacillus subtilis: regulation and involvement in the biosyntheses of menaquinone and 2,3-dihydroxybenzoate

Rowland

Taber

1996

J Bacteriol

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Bacillus subtilis has duplicate isochorismate synthase genes, menF and dhbC. Isochorismate synthase is involved in the biosynthesis of both the respiratory chain component menaquinone (MK) and the siderophore 2,3-dihydroxybenzoate (DHB). Several menF and dhbC deletion mutants were constructed to identify the contribution made by each gene product to MK and DHB biosynthesis. menF deletion mutants were able to produce wild-type levels of MK and DHB, suggesting that the dhbC gene product is able to compensate for the lack of MenF. However, a dhbC deletion mutant produced wild-type levels of MK but was DHB deficient, indicating that MenF is unable to compensate for the lack of DhbC. A menF dhbC double-deletion mutant was both MK and DHB deficient. Transcription analysis showed that expression of dhbC, but not of menF, is regulated by iron concentration. A dhbA::lacZ fusion strain was constructed to examine the effects of mutations to the iron box sequence within the dhb promoter region. These mutations abolished the iron-regulated transcription of the dhb genes, suggesting that a Fur-like repressor protein exists in B. subtilis.Isochorismate synthase is responsible for converting chorismate to isochorismate and is necessary for the biosynthesis of both the respiratory chain component menaquinone (MK) and the Bacillus subtilis siderophore 2,3-dihydroxybenzoate (DHB) (Fig. 1). Although it is known that the carbon source, growth phase, and oxygen tension have regulatory effects, the actual signals that induce expression of respiratory chain components are unknown (37). However, the production of DHB is known to be regulated by iron concentration (1,25,26). Therefore, isochorismate is required by cells under very different environmental conditions.B. subtilis has two distinct isochorismate synthases encoded by the menF (30) and dhbC (29) genes. MenF and DhbC are 47% identical at the DNA level and have 35% amino acid identity (29). menF is a promoter-proximal gene of the MK biosynthetic gene cluster located at 273Њ on the chromosome (30,38). MK is a lipophilic, nonprotein redox component mediating electron transfer between dehydrogenases and cytochromes (37). dhbC is the second gene of the DHB biosynthetic gene cluster located at 291Њ on the chromosome (29). Under conditions of iron deprivation, B. subtilis synthesizes DHB, a component of the specific transport system for the uptake of extracellular iron (1,25,26). This is not the only instance of gene duplication in B. subtilis. There are two thymidylate synthetases, TSaseA and TSaseB, that are encoded by the unlinked thyA and thyB genes, respectively (22). Both enzymes are functional in bacteria grown at 37ЊC or lower temperatures, but only TSaseA is active at 46ЊC (22). There are two different a-type terminal oxidases in B. subtilis that are encoded by separate loci (32). One of the oxidases is associated with a heme C-containing unit (32). The two membrane alkaline phosphatases of B. subtilis have substantial differences with respect to molecular weight, substrate sp...

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“…First, when the intact ctaB-ctaC intercistronic sequence was cloned in front of promoterless lacZ at the amyE locus, as either an in-frame translational or a transcriptional fusion, no ␤-galactosidase activity could be detected (data not shown). However, the same translational construct has been successfully used as an expression indicator for menp 1 , the major promoter in menaquinone biosynthesis (32). Second, when primer extension and rapid amplification of 5Ј cDNA ends were carried out with primers complementary to the 5Ј region of the ctaC open reading frame, no ctaCЈ transcriptional start sites were found.…”

Section: Resultsmentioning

confidence: 99%

Catabolite Regulation of the Bacillus subtilis ctaBCDEF Gene Cluster

Liu

Taber

1998

J Bacteriol

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Bacillus subtilis cytochrome c oxidase caa 3 is encoded by the ctaCDEF genes at the ctaABCDEF locus, with the ctaBCDEF genes organized as an operon-like unit. A dyad symmetry sequence and a catabolite response element homolog can be recognized in the 240-bp intercistronic region between ctaB and ctaC. ctaB-lacZ and ctaBCD-lacZ transcriptional fusions integrated at the native locus were used to study catabolite effects on transcription of the ctaB and ctaCDEF genes. In Schaeffer's medium lacking glucose, ctaBCD-lacZ was expressed at a very low level during the exponential phase, and expression increased about 30-fold 2 h after entry into the stationary phase. In the presence of 0.5% glucose, ctaBCD-lacZ expression was totally repressed. In contrast to ctaBCD-lacZ, ctaB-lacZ was constitutively expressed regardless of carbon source. The ctaCDEF genes were separated from ctaB by insertion of plasmids carrying selectable markers in such a way that the ctaCDEF and ctaB transcription units remained intact. Enzymatic assays of caa 3 with these constructs, showed that ctaCDEF was not expressed independently of ctaB. Also, when a ctaB-ctaC-lacZ fusion (containing the ctaB-ctaC intercistronic region) was placed at a remote nonessential locus, ␤-galactosidase activity could not be detected. The absence of a promoter in the ctaB-ctaC intercistronic space also was indicated by the inability to detect ctaC-specific transcripts with RNase protection assays, primer extension, and rapid amplification of 5 cDNA ends. Direct mRNA measurements showed that, in the presence of 0.5% glucose, ctaBCDEF transcripts terminated at the 3 end of the putative stem-loop structure and the distal portion was down-regulated. A possible mechanism for ctaCDEF gene regulation is suggested. Catabolite repression of ctaBCD-lacZ was partly dependent on CcpA but was independent of HPr. The expression of ctaBCDEF also appears to require the strC, ctaA, and resD-resE gene products.

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Transcriptional regulation of the Bacillus subtilis menp1 promoter

Cited by 10 publications

References 32 publications

Phenotype Microarray Profiling of Staphylococcus aureus menD and hemB Mutants with the Small-Colony-Variant Phenotype

Phenotype Microarray Profiling of Staphylococcus aureus menD and hemB Mutants with the Small-Colony-Variant Phenotype

Duplicate isochorismate synthase genes of Bacillus subtilis: regulation and involvement in the biosyntheses of menaquinone and 2,3-dihydroxybenzoate

Catabolite Regulation of the Bacillus subtilis ctaBCDEF Gene Cluster

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