Identification of a Catabolite-Responsive Element Necessary for Regulation of the
 cry4A
 Gene of
 Bacillus thuringiensis
 subsp.
 israelensis

Kant, Sashi; Kapoor, Rupam; Banerjee, Nirupama

doi:10.1128/jb.00421-09

Cited by 8 publications

(9 citation statements)

References 33 publications

(34 reference statements)

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“…The 363-bp upstream region preceding the parD2 start codon (P 363parD2 ) of M. tuberculosis H37Rv was PCR-amplified and fused to a promoter-less lacZ reporter gene (3,067 bp) in pSK6 plasmid ( Kant et al, 2009 ). The 363-bp lacZ fusion fragment was subsequently cloned in an E. coli - Mycobacterium shuttle vector pMV261, using NotI and DraI sites, forming pMS2 (Supplementary Figure S2 ).…”

Section: Methodsmentioning

confidence: 99%

The Chromosomal parDE2 Toxin–Antitoxin System of Mycobacterium tuberculosis H37Rv: Genetic and Functional Characterization

et al. 2016

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Mycobacterium tuberculosis H37Rv escapes host-generated stresses by entering a dormant persistent state. Activation of toxin-antitoxin modules is one of the mechanisms known to trigger such a state with low metabolic activity. M. tuberculosis harbors a large number of TA systems mostly located within discernible genomic islands. We have investigated the parDE2 operon of M. tuberculosis H37Rv encoding MParE2 toxin and MParD2 antitoxin proteins. The parDE2 locus was transcriptionally active from growth phase till late stationary phase in M. tuberculosis. A functional promoter located upstream of parD2 GTG start-site was identified by 5′-RACE and lacZ reporter assay. The MParD2 protein transcriptionally regulated the PparDE2 promoter by interacting through Arg16 and Ser15 residues located in the N-terminus. In Escherichia coli, ectopic expression of MParE2 inhibited growth in early stages, with a drastic reduction in colony forming units. Live-dead analysis revealed that the reduction was not due to cell death alone but due to formation of viable but non-culturable cells (VBNCs) also. The toxic activity of the protein, identified in the C-terminal residues Glu98 and Arg102, was neutralized by the antitoxin MParD2, both in vivo and in vitro. MParE2 inhibited mycobacterial DNA gyrase and interacted with the GyrB subunit without affecting its ATPase activity. Introduction of parE2 gene in the heterologous M. smegmatis host prevented growth and colony formation by the transformed cells. An M. smegmatis strain containing the parDE2 operon also switched to a non-culturable phenotype in response to oxidative stress. Loss in colony-forming ability of a major part of the MParE2 expressing cells suggests its potential role in dormancy, a cellular strategy for adaptation to environmental stresses. Our study has laid the foundation for future investigations to explore the physiological significance of parDE2 operon in mycobacterial pathogenesis.

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Section: Methodsmentioning

confidence: 99%

The Chromosomal parDE2 Toxin–Antitoxin System of Mycobacterium tuberculosis H37Rv: Genetic and Functional Characterization

et al. 2016

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“…It is known that a complex built of cAMP and its receptor protein (CRP) binds to these recognition sequences. By using a consensus sequence for CRE sites ( Kant et al, 2009 ), the 5′ untranslated region (5′-UTR) of the sodorifen cluster was screened and two possible binding sites of the cAMP/CRP complex (CRE1/2) with an identity of 85.7 and 78.6%, respectively, to the consensus sequence were found ( Figure 2 ). Furthermore, the potential CRE sites identified in S.p.…”

Section: Resultsmentioning

confidence: 99%

“…Potential CRE binding sites were identified with the help of the Regulatory Sequence Analysis Tool for prokaryotes (RSAT; Medina-Rivera et al, 2015 ) using an already published CRE consensus sequence ( Kant et al, 2009 ) as a motif to search in the 5′ UTR of the S.p. 4Rx13 sodorifen cluster.…”

Section: Methodsmentioning

confidence: 99%

Carbon Catabolite Repression Regulates the Production of the Unique Volatile Sodorifen of Serratia plymuthica 4Rx13

2017

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Microorganisms are capable of synthesizing a plethora of secondary metabolites including the long-overlooked volatile organic compounds. Little knowledge has been accumulated regarding the regulation of the biosynthesis of such mVOCs. The emission of the unique compound sodorifen of Serratia plymuthica isolates was significantly reduced in minimal medium with glucose, while succinate elevated sodorifen release. The hypothesis of carbon catabolite repression (CCR) acting as a major control entity on the synthesis of mVOCs was proven by genetic evidence. Central components of the typical CCR of Gram-negative bacteria such as the adenylate cyclase (CYA), the cAMP binding receptor protein (CRP), and the catabolite responsive element (CRE) were removed by insertional mutagenesis. CYA, CRP, CRE1 mutants revealed a lower sodorifen release. Moreover, the emission potential of other S. plymuthica isolates was also evaluated.

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“…In Gram-positive bacteria, catabolite repression of many catabolic operons involves the phosphocarrier protein HPr, the catabolite control protein CcpA, and a cis -acting catabolite responsive element ( cre ) [ 13 , 14 ]. The phosphorylated HPr binds to the CcpA to form a complex with strong DNA binding affinity [ 47 , 48 ].…”

Section: Metabolic Regulation Of Cry Protein Productionmentioning

confidence: 99%

“…The phosphorylated HPr-CcpA complex modulates the transcription of target genes by binding to the cre sequence [ 49 ]. Glucose represses cry4A gene expression at the mRNA level in Bti [ 50 ], and the phosphorylated HPr-CcpA complex of Bti represses cry4A transcription by specifically binding to a 15 bp cre sequence overlapping the -35 element of the cry4A promoter [ 14 ]. Glucose catabolite repression of cry4A is abolished both by site-specific mutation of the cre sequence [ 14 ] and by the HPr-S45A mutant, which produces phosphorylation-disabled HPr [ 13 ]: both increase the activity of the cry4A promoter.…”

Section: Metabolic Regulation Of Cry Protein Productionmentioning

confidence: 99%

Regulation of cry Gene Expression in Bacillus thuringiensis

Deng

Peng

Song

et al. 2014

Toxins

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Bacillus thuringiensis differs from the closely related Bacillus cereus group species by its ability to produce crystalline inclusions. The production of these crystals mainly results from the expression of the cry genes, from the stability of their transcripts and from the synthesis, accumulation and crystallization of large amounts of insecticidal Cry proteins. This process normally coincides with sporulation and is regulated by various factors operating at the transcriptional, post-transcriptional, metabolic and post-translational levels.

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Identification of a Catabolite-Responsive Element Necessary for Regulation of the cry4A Gene of Bacillus thuringiensis subsp. israelensis

Cited by 8 publications

References 33 publications

The Chromosomal parDE2 Toxin–Antitoxin System of Mycobacterium tuberculosis H37Rv: Genetic and Functional Characterization

The Chromosomal parDE2 Toxin–Antitoxin System of Mycobacterium tuberculosis H37Rv: Genetic and Functional Characterization

Carbon Catabolite Repression Regulates the Production of the Unique Volatile Sodorifen of Serratia plymuthica 4Rx13

Regulation of cry Gene Expression in Bacillus thuringiensis

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