title: A DNA adenine methyltransferase of Escherichia coli that is cell cycle regulated and essential for viability.
abstract: DNA sequence analysis revealed that the putative yhdJ DNA methyltransferase gene of Escherichia coli is 55% identical to the Nostoc sp. strain PCC7120 gene encoding DNA methyltransferase AvaIII, which methylates adenine in the recognition sequence, ATGCAT. The yhdJ gene was cloned, and the enzyme was overexpressed and purified. Methylation and restriction analysis showed that the DNA methyltransferase methylates the first adenine in the sequence ATGCAT. This DNA methylation was found to be regulated during the cell cycle, and the DNA adenine methyltransferase was designated M.EcoKCcrM (for "cell cycle-regulated methyltransferase"). The CcrM DNA adenine methyltransferase is required for viability in E. coli, as a strain lacking a functional genomic copy of ccrM can be isolated only in the presence of an additional copy of ccrM supplied in trans. The cells of such a knockout strain stopped growing when expression of the inducible plasmid ccrM gene was shut off. Overexpression of M.EcoKCcrM slowed bacterial growth, and the ATGCAT sites became fully methylated throughout the cell cycle; a high proportion of cells with an anomalous size distribution and DNA content was found in this population. Thus, the temporal control of this methyltransferase may contribute to accurate cell cycle control of cell division and cellular morphology. Homologs of M.EcoKCcrM are present in other bacteria belonging to the gamma subdivision of the class Proteobacteria, suggesting that methylation at ATGCAT sites may have similar functions in other members of this group.
authors.affiliation: Sealy Center for Molecular Science and Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch at Galveston, Galveston, Texas 77555-1071, USA. vakossyk@utmb.edu
authors.forename: Valeri G
authors.initials: VG
authors.lastname: Kossykh
authors.forename: R Stephen
authors.initials: RS
authors.lastname: Lloyd
mesh_terms: D000595:Amino Acid Sequence; D002455:Cell Division; D003470:Culture Media; D019175:DNA Methylation; D004926:Escherichia coli / Q000166:cytology* / Q000201:enzymology; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000378:metabolism*
chemical_list: D003470:Culture Media; C098075:DNA modification methylase CcrM; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)