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Title

DNA binding of the cell cycle transcriptional regulator GcrA depends on N6-adenosine methylation in Caulobacter crescentus and other Alphaproteobacteria

Authors
Fioravanti, Antonella
Mohapatra, Saswat S
Bompard, Coralie
Brilli, Matteo
Castric, Vincent
Villeret, Vincent
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Published in PLOS Genetics. 2013, vol. 9, no. 5, p. e1003541
Abstract Several regulators are involved in the control of cell cycle progression in the bacterial model system Caulobacter crescentus, which divides asymmetrically into a vegetative G1-phase (swarmer) cell and a replicative S-phase (stalked) cell. Here we report a novel functional interaction between the enigmatic cell cycle regulator GcrA and the N6-adenosine methyltransferase CcrM, both highly conserved proteins among Alphaproteobacteria, that are activated early and at the end of S-phase, respectively. As no direct biochemical and regulatory relationship between GcrA and CcrM were known, we used a combination of ChIP (chromatin-immunoprecipitation), biochemical and biophysical experimentation, and genetics to show that GcrA is a dimeric DNA-binding protein that preferentially targets promoters harbouring CcrM methylation sites. After tracing CcrM-dependent N6-methyl-adenosine promoter marks at a genome-wide scale, we show that these marks recruit GcrA in vitro and in vivo. Moreover, we found that, in the presence of a methylated target, GcrA recruits the RNA polymerase to the promoter, consistent with its role in transcriptional activation. Since methylation-dependent DNA binding is also observed with GcrA orthologs from other Alphaproteobacteria, we conclude that GcrA is the founding member of a new and conserved class of transcriptional regulators that function as molecular effectors of a methylation-dependent (non-heritable) epigenetic switch that regulates gene expression during the cell cycle.
Keywords Adenosine/geneticsAlphaproteobacteria/growth & developmentAmino Acid SequenceCaulobacter crescentus/genetics/growth & developmentCell Cycle Proteins/genetics/metabolismDNA Methylation/geneticsDNA-Binding Proteins/geneticsEpigenesis, GeneticGene Expression Regulation, BacterialMethyltransferases/genetics/metabolismPromoter Regions, GeneticTranscription, Genetic
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PMID: 23737758
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Article (Published version) (7 MB) - public document Free access
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Research group Régulation du cycle cellulaire dans C.crescentus, une bactérie asymétrique (895)
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FIORAVANTI, Antonella et al. DNA binding of the cell cycle transcriptional regulator GcrA depends on N6-adenosine methylation in Caulobacter crescentus and other Alphaproteobacteria. In: PLOS Genetics, 2013, vol. 9, n° 5, p. e1003541. https://archive-ouverte.unige.ch/unige:32604

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Deposited on : 2013-12-20

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