Genome-wide analysis of deoxyadenosine methyltransferase-mediated control of gene expression in Escherichia coli.
Deoxyadenosine methyltransferase (Dam) methylates the deoxyadenine residues in 5'-GATC-3' sequences and is important in many cellular processes in Escherichia coli. We performed a computational analysis of the entire E. coli genome and confirmed that GATC sequences are distributed unevenly in regulatory regions, which suggests that Dam might regulate gene transcription. To test this, a high-density DNA microarray of 4097 E. coli genes was constructed and used to assess the gene expression profiles of the wild type and the dam-16::kam mutant strain grown under four different conditions. We also used two-dimensional electrophoretic analysis of the proteome to assess the protein profiles. The expression of a large number of genes was affected by the dam deficiency. Genes involved in aerobic respiration, stress and SOS responses, amino acid metabolism and nucleotide metabolism were expressed at higher levels in the mutant cells, especially in aerobic conditions. In contrast, transcription of genes participating in anaerobic respiration, flagella biosynthesis, chemotaxis and motility was decreased in the dam mutant strain under both aerobic and low aerobic conditions. Thus, Dam-controlled genes are involved in adjusting the metabolic and respiratory pathways and bacterial motility to suit particular environmental conditions. The promoters of most of these Dam-controlled genes were also found to contain GATC sequences that overlap with recognition sites for two global regulators, fumarate nitrate reduction (Fnr) and catabolite activator protein (CRP). We propose that Dam-mediated methylation plays an important role in the global regulation of genes, particularly those with Fnr and CRP binding sites.
CREST, JST (Japan Science and Technology).
Taku
T
Oshima
Chieko
C
Wada
Yuya
Y
Kawagoe
Takeshi
T
Ara
Maki
M
Maeda
Yasushi
Y
Masuda
Sota
S
Hiraga
Hirotada
H
Mori
D000596:Amino Acids / Q000378:metabolism; D015152:Blotting, Northern; D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D007506:Iron-Sulfur Proteins; D009711:Nucleotides / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D033902:Periplasmic Binding Proteins / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D012486:Salmonella typhimurium / Q000235:genetics; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D000596:Amino Acids; D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D029968:Escherichia coli Proteins; C060704:FNR protein, E coli; D007506:Iron-Sulfur Proteins; D009711:Nucleotides; D033902:Periplasmic Binding Proteins; D010743:Phospholipids; C058693:Dam methyltransferase; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); C499294:dam protein, E coli