<scp>ComE</scp>/<scp>ComE</scp>∼<scp>P</scp> interplay dictates activation or extinction status of pneumococcal <scp>X</scp>‐state (competence)

Martin, Bernard; Soulet, Anne-Lise; Mirouze, Nicolas; Prudhomme, Marc; Mortier‐Barrière, Isabelle; Granadel, Chantal; Noirot‐Gros, Marie‐Françoise; Naveilhan, Philippe; Polard, Patrice; Claverys, Jean‐Pierre

doi:10.1111/mmi.12104

Cited by 89 publications

(163 citation statements)

References 44 publications

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“…The 'open' form of the ComE D58A -comcde complex that we observed experimentally may be the intermediate suggested by Martin et al [10]. These authors proposed that the extinction of the competence state follows the breakdown of the REC dimer, which is caused by interaction with DprA.…”

Section: D58esupporting

confidence: 64%

“…ComD senses the extracellular concentration of CSP [3], leading to autophosphorylation on H248, and the phosphoryl group is then transferred to residue D58 of ComE. The activated ComE binds two imperfect direct repeat motifs, DR1 and DR2 (9 bp each, separated by a 12-mer linker), in the promoter regions and induces the expression of the comAB, comCDE and comX operons [7][8][9][10]. The comAB operon encodes the machinery required for the maturation and export of pre-CSP, whereas activation of the comCDE operon creates an auto-catalytic, rapid and synchronous activation of competence [6].…”

Section: Introductionmentioning

confidence: 99%

“…ComE D58A abolishes basal comCDE expression. In contrast, DcomD S. pneumoniae cells expressing ComE D58E display full spontaneous competence [10,20]. Using these two D58A and D58E mutants, we have recently provided a structural study of ComE, which is composed of two domains: REC (the regulation domain via the phosphorylation of D58) and LytTR (the effector domain) [22].…”

Section: Introductionmentioning

confidence: 99%

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Modeling the ComD/ComE/comcde interaction network using small angle X‐ray scattering

et al. 2015

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The ComD–ComE two‐component system controls the competence state of Streptococcus pneumoniae via the phospho‐regulation of ComE, which fluctuates between monomeric and dimeric states. We previously showed that the non‐phosphorylatable ComED58A mutant is monomeric in solution, whereas the ComED58E active mimic mutant dimerizes via its REC domains. The crystal structure of ComED58A revealed an asymmetric dimer that may represent the activated form of ComE. Here, we investigated the binding between the catalytic domain of ComD, ComE and the promoter region comcde, using small angle X‐ray scattering. ComDcatdom is a dimer that adapts two monomers of ComE, one on each side, placing ComED58 residue in front of ComDH248, a location that is convenient for the intermolecular transfer reaction of the phosphoryl group. The LytTR, ComED58A and ComED58E complexed with comcde are composed of two protein molecules per DNA duplex. Modeling the complexes against small angle X‐ray scattering data indicated that ComED58E bound to comcde forms a compact dimer similar to the crystal structure, whereas ComED58A–comcde adopts more than one conformation with or without dimer contacts. The various oligomeric states of ComE induce different bending angles of the promoter, which provides a mechanistic scenario for the activation of ComE: the phosphorylation of ComE forces additional bending of comcde, and the release of this bending strain on DNA via the disruption of the ComE dimer may signal the shut‐off of the competence state. Database The molecular models and experimental SAXS data have been deposited on SASBDB (Small Angle Scattering Biological Data Bank) (see http://www.sasbdb.org/aboutSASBDB/) under the SAS codes SASDAA7, SASDAB7 and SASDAC7.

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Section: D58esupporting

confidence: 64%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Modeling the ComD/ComE/comcde interaction network using small angle X‐ray scattering

et al. 2015

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“…Cells in the exponential growth phase are first primed for transformation by a QS mechanism encoded by two genetic loci initially transcribed at a basal level by core RNA polymerase (RNAP) bound to the primary sigma factor, A (3). The loci are comAB (4,5) and comCDE (3).…”

mentioning

confidence: 99%

“…The loci are comAB (4,5) and comCDE (3). ComC is a propeptide cleaved and exported by ComAB, as the mature peptide called CSP (competence-stimulating peptide) (6).…”

mentioning

confidence: 99%

Competence for Genetic Transformation in Streptococcus pneumoniae: Mutations in σ ^A Bypass the ComW Requirement for Late Gene Expression

2016

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Streptococcus pneumoniae is able to integrate exogenous DNA into its genome by natural genetic transformation. Transient accumulation of high levels of the only S. pneumoniae alternative factor is insufficient for development of full competence without expression of a second competence-specific protein, ComW. The ⌬comW mutant is 10 4 -fold deficient in the yield of recombinants, 10-fold deficient in the amount of X activity, and 10-fold deficient in the amount of X protein. The critical role of ComW during transformation can be partially obviated by A mutations clustered on surfaces controlling affinity for core RNA polymerase (RNAP). While strains harboring A mutations in the comW mutant background were transforming at higher rates, the mechanism of transformation restoration was not clear. To investigate the mechanism of transformation restoration, we measured late gene expression in A * suppressor strains. Restoration of late gene expression was observed in ⌬comW A * mutants, indicating that a consequence of the A * mutations is, at least, to restore X activity. Competence kinetics were normal in ⌬comW A * strains, indicating that strains with restored competence exhibit the same pattern of transience as wild-type (WT) strains. We also identified a direct interaction between ComW and X using the yeast two-hybrid (Y2H) assay. Taken together, these data are consistent with the idea that ComW increases X access to core RNAP, pointing to a direct role of ComW in factor exchange during genetic transformation. However, the lack of late gene shutoff in ⌬comW mutants also points to a potential new role for ComW in competence shutoff. IMPORTANCEThe sole alternative sigma factor of the streptococci, SigX, regulates development of competence for genetic transformation, a widespread mechanism of adaptation by horizontal gene transfer in this genus. The transient appearance of this sigma factor is strictly controlled at the levels of transcription and stability. This report shows that it is also controlled at the point of its substitution for SigA by a second transient competence-specific protein, ComW. Streptococcus pneumoniae, a Gram-positive opportunistic pathogen found in the human nasopharynx, causes diseases such as pneumonia and meningitis. S. pneumoniae's natural competence, or ability to integrate exogenous DNA into its chromosome, provides a major mechanism of rapidly overcoming selective pressure (1, 2). The ability to take up and exchange DNA depends on development of the competent state, which is prompted by quorum sensing (QS) mediated by a small peptide, via a signal transduction pathway that is incompletely understood.Cells in the exponential growth phase are first primed for transformation by a QS mechanism encoded by two genetic loci initially transcribed at a basal level by core RNA polymerase (RNAP) bound to the primary sigma factor, A (3). The loci are comAB (4, 5) and comCDE (3). ComC is a propeptide cleaved and exported by ComAB, as the mature peptide called CSP (competence-stimulating pepti...

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Clp ATPases differentially affect natural competence development in Streptococcus mutans

Pandey

Biswas

2022

MicrobiologyOpen

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In naturally competent bacteria, DNA transformation through horizontal gene transfer is an evolutionary mechanism to receive extracellular DNA. Bacteria need to maintain a state of competence to accept foreign DNA, and this is an energy-driven phenomenon that is tightly controlled. In Streptococcus, competence development is a complex process that is not fully understood. In this study, we used Streptococcus mutans, an oral bacterium, to determine how cell density affects competence development. We found that in S. mutans the transformation efficiency is maximum when the transforming DNA was added at low cell density and incubated for 2.5 h before selecting for transformants. We also found that S. mutans cells remain competent until the mid-logarithmic phase, after which the competence decreases drastically. Surprisingly, we observed that individual components of Clp proteolytic complexes differentially regulate competence. If the transformation is carried out at the early growth phase, both ClpP protease and ClpX ATPase are needed for competence. In contrast, we found that both ClpC and ClpE negatively affect competence. We also found that if the transformation is carried out at the mid-logarithmic growth phase ClpX is still required for competence, but ClpP negatively affects competence. While the exact reason for this differential effect of ClpP and ClpX on transformation is currently unknown, we found that both ClpC and ClpE have a negative effect on transformation, which was not reported before.

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ComE/ComE∼P interplay dictates activation or extinction status of pneumococcal X‐state (competence)

Cited by 89 publications

References 44 publications

Modeling the ComD/ComE/comcde interaction network using small angle X‐ray scattering

Modeling the ComD/ComE/comcde interaction network using small angle X‐ray scattering

Competence for Genetic Transformation in Streptococcus pneumoniae: Mutations in σ ^A Bypass the ComW Requirement for Late Gene Expression

Clp ATPases differentially affect natural competence development in Streptococcus mutans

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ComE/ComE∼P interplay dictates activation or extinction status of pneumococcal X‐state (competence)

Cited by 89 publications

References 44 publications

Modeling the ComD/ComE/comcde interaction network using small angle X‐ray scattering

Modeling the ComD/ComE/comcde interaction network using small angle X‐ray scattering

Competence for Genetic Transformation in Streptococcus pneumoniae: Mutations in σ A Bypass the ComW Requirement for Late Gene Expression

Clp ATPases differentially affect natural competence development in Streptococcus mutans

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Competence for Genetic Transformation in Streptococcus pneumoniae: Mutations in σ ^A Bypass the ComW Requirement for Late Gene Expression