title: The functions of DNA methylation by CcrM in Caulobacter crescentus: a global approach.
abstract: DNA methylation is involved in a diversity of processes in bacteria, including maintenance of genome integrity and regulation of gene expression. Here, using Caulobacter crescentus as a model, we exploit genome-wide experimental methods to uncover the functions of CcrM, a DNA methyltransferase conserved in most Alphaproteobacteria. Using single molecule sequencing, we provide evidence that most CcrM target motifs (GANTC) switch from a fully methylated to a hemi-methylated state when they are replicated, and back to a fully methylated state at the onset of cell division. We show that DNA methylation by CcrM is not required for the control of the initiation of chromosome replication or for DNA mismatch repair. By contrast, our transcriptome analysis shows that >10% of the genes are misexpressed in cells lacking or constitutively over-expressing CcrM. Strikingly, GANTC methylation is needed for the efficient transcription of dozens of genes that are essential for cell cycle progression, in particular for DNA metabolism and cell division. Many of them are controlled by promoters methylated by CcrM and co-regulated by other global cell cycle regulators, demonstrating an extensive cross talk between DNA methylation and the complex regulatory network that controls the cell cycle of C. crescentus and, presumably, of many other Alphaproteobacteria.
authors.affiliation: Department of Fundamental Microbiology, University of Lausanne, Lausanne, CH 1015, Switzerland, Department of Developmental Biology, Stanford University, CA 94305, USA and Department of Chemistry, Stanford University, CA 94305, USA.
authors.forename: Diego
authors.initials: D
authors.lastname: Gonzalez
authors.forename: Jennifer B
authors.initials: JB
authors.lastname: Kozdon
authors.forename: Harley H
authors.initials: HH
authors.lastname: McAdams
authors.forename: Lucy
authors.initials: L
authors.lastname: Shapiro
authors.forename: Justine
authors.initials: J
authors.lastname: Collier
mesh_terms: D020561:Alphaproteobacteria / Q000235:genetics; D001426:Bacterial Proteins / Q000145:classification / Q000378:metabolism*; D016935:Caulobacter crescentus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D002876:Chromosomes, Bacterial; D019175:DNA Methylation; D015964:Gene Expression Regulation, Bacterial; D016680:Genome, Bacterial; D059645:Mutation Rate; D059372:Nucleotide Motifs; D010802:Phylogeny; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000145:classification / Q000378:metabolism*; D059467:Transcriptome
chemical_list: D001426:Bacterial Proteins; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)