Substitutional Analysis of the C-Terminal Domain of AbrB Revealed Its Essential Role in DNA-Binding Activity

Neubauer, Svetlana; Dolgova, Olga; Präg, Gregory; Borriss, Rainer; Makarewicz, Oliwia

doi:10.1371/journal.pone.0097254

Cited by 6 publications

(5 citation statements)

References 41 publications

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“… B. licheniformis M18 displays a deletion in abrB , affecting extracellular enzyme secretion and natural competence development. (a) The abrB genomic region of B. licheniformis 9945A (WT), its derivative M18 and the corresponding amino acid sequences with domain indications (Neubauer et al , 2014; Phillips & Strauch, 2001; Xu et al , 1996). ORFs are depicted as arrows; dashed lines refer to the deletion in M18.…”

Section: Resultsmentioning

confidence: 99%

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Unravelling the genetic basis for competence development of auxotrophic Bacillus licheniformis 9945A strains

et al. 2014

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Bacterial natural genetic competence -well studied in Bacillus subtilis -enables cells to take up and integrate extracellularly supplied DNA into their own genome. However, little is known about competence development and its regulation in other members of the genus, although DNA uptake machineries are routinely encoded. Auxotrophic Bacillus licheniformis 9945A derivatives, obtained from repeated rounds of random mutagenesis, were long known to develop natural competence. Inspection of the colony morphology and extracellular enzyme secretion of two of these derivatives, M28 and M18, suggested that regulator genes are collaterally hit. M28 emerged as a 14 bp deletion mutant concomitantly displaying a shift in the reading frame of degS that encodes the sensor histidine kinase, which is part of the molecular switch that directs cells to genetic competence, the synthesis of extracellular enzymes or biofilm formation, while for M18, sequencing of the suspected gene revealed a 375 bp deletion in abrB, encoding the major transition state regulator. With respect to colony morphology, enzyme secretion and competence development, both of the mutations, when newly generated on the wild-type B. licheniformis 9945A genetic background, resulted in phenotypes resembling M28 and M18, respectively. All of the known naturally competent B. licheniformis representatives, hitherto thoroughly investigated in this regard, carry mutations in regulator genes, and hence genetic competence observed in domesticated strains supposedly results from deregulation.

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Section: Resultsmentioning

confidence: 99%

“…WT, wild-type; PGA", polyglutamate deficient; Gly", glycine auxotroph. (Neubauer et al, 2014;Phillips & Strauch, 2001;Xu et al, 1996). ORFs are depicted as arrows; dashed lines refer to the deletion in M18.…”

Section: M28 Reveals a Frame Shift In Degsmentioning

confidence: 99%

Unravelling the genetic basis for competence development of auxotrophic Bacillus licheniformis 9945A strains

et al. 2014

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“…S5). Previous studies show that both the N-terminus and C-terminus of AbrB from B. subtilis are essential for its function (Neubauer et al, 2014;Olson et al, 2014); however, the critical residues involved remain unknown. Sequence comparison of the three AbrB proteins studied in this work may provide important information on the residues required for regulation of activity and/or sequencespecific DNA binding.…”

Section: Discussionmentioning

confidence: 99%

“…2A). In fact, both CcpA and AbrB have previously been found to be global transcription factors, with CcpA functioning to negatively regulate many pathways involved in carbon source utilization in Gram-positive bacteria (Warner and Lolkema, 2003), while AbrB functioning as a transition state regulator to extensively repress gene expression during the exponential growth in B. subtilis (Neubauer et al, 2014). Given the positions of the putative binding sites of CcpA and AbrB to be located in the 5 0 -UTR and to overlap with the −35 box of the prp promoter, respectively ( Fig.…”

Section: Ccpa and Abrb Repress Transcription Of The Prp Operonmentioning

confidence: 99%

2‐Methylcitrate cycle: a well‐regulated controller of Bacillus sporulation

Cao

Du³

et al. 2019

Environmental Microbiology

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Summary Bacillus thuringiensis is the most widely used eco‐friendly biopesticide, containing two primary determinants of biocontrol, endospore and insecticidal crystal proteins (ICPs). The 2‐methylcitrate cycle is a widespread carbon metabolic pathway playing a crucial role in channelling propionyl‐CoA, but with poorly understood metabolic regulatory mechanisms. Here, we dissect the transcriptional regulation of the 2‐methylcitrate cycle operon prpCDB and report its unprecedented role in controlling the sporulation process of B. thuringiensis. We found that the transcriptional activity of the prp operon encoding the three critical enzymes PrpC, PrpD, and PrpB in the 2‐methylcitrate cycle was negatively regulated by the two global transcription factors CcpA and AbrB, while positively regulated by the LysR family regulator CcpC, which jointly account for the fact that the 2‐methylcitrate cycle is specifically and highly active in the stationary phase of growth. We also found that the prpD mutant accumulated 2‐methylcitrate, the intermediate metabolite of the 2‐methylcitrate cycle, which delayed and inhibited sporulation at the early stage. Thus, our results not only revealed sophisticated transcriptional regulatory mechanisms for the metabolic 2‐methylcitrate cycle but also identified 2‐methylcitrate as a novel regulator of sporulation in B. thuringiensis.

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“…AbrBC mutations at N64 and L67 are also known to affect the multimerization state and hence DNA binding (Strauch – unpublished data). AbrBC mutations at Q55, K71, E80, Q81, E85 and E90 have also been proposed to affect DNA-binding (Neubauer et al, 2014). Since no controls showing wild-type AbrBC binding to DNA are available at this time, it is not possible to fully discern whether DNA-binding is impeded due to direct binding or by changes in charge packing/multimerization of AbrBC.…”

Section: Discussionmentioning

confidence: 99%

Structure and DNA-Binding Traits of the Transition State Regulator AbrB

et al. 2014

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Summary The AbrB protein from Bacillus subtilis is a DNA-binding global regulator controlling the onset of a vast array of protective functions under stressful conditions. Such functions include biofilm formation, antibiotic production, competence development, extracellular enzyme production, motility and sporulation. AbrB orthologues are known in a variety of prokaryotic organisms, most notably in all infectious strains of Clostridia, Listeria and Bacilli. Despite its central role in bacterial response and defense, its structure has been elusive, due to its highly dynamic character. Orienting its N- and C-terminal domains with respect to one another has been especially problematic. Here we have generated the first structure of full-length, tetrameric AbrB using NMR, chemical crosslinking and mass spectrometry. We note that AbrB possesses a strip of positive electrostatic potential encompassing its DNA binding region and that its C-terminal domain aids in DNA binding.

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Substitutional Analysis of the C-Terminal Domain of AbrB Revealed Its Essential Role in DNA-Binding Activity

Cited by 6 publications

References 41 publications

Unravelling the genetic basis for competence development of auxotrophic Bacillus licheniformis 9945A strains

Unravelling the genetic basis for competence development of auxotrophic Bacillus licheniformis 9945A strains

2‐Methylcitrate cycle: a well‐regulated controller of Bacillus sporulation

Structure and DNA-Binding Traits of the Transition State Regulator AbrB

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