Nucleotide sequence and expression of the Escherichia coli dapB gene.
The Escherichia coli dapB gene encodes dihydrodipicolinate reductase. This enzyme is part of the diaminopimelate-lysine pathway, and its synthesis is repressed by lysine. The dapB gene was cloned into pBR322 from a transducing lambda bacteriophage, its complete nucleotide sequence established, and the transcriptional start localized. The DNA sequence predicts that the dapB gene codes for a 273-amino acid polypeptide, Mr 28,798. No attenuation-type sequence can be found between the mRNA start and the coding sequence. The dapB promoter signals appear to be weak as compared to RNA polymerase consensus sequences. Nevertheless an efficient in vivo synthesis of beta-galactosidase was obtained when the lac operon was inserted in vitro in the dapB gene, downstream of the dapB regulatory signals. Further studies were performed on an in-frame gene fusion constructed in vitro between the dapB and the lacZ genes. They indicated that repression by lysine is exerted on a DNA region restricted to a 153-base pair fragment with only 102 nontranscribed nucleotides. Finally, dapB gene expression showed a gene dosage effect which suggests that it is not controlled by an element present in limiting amounts in the cell.
D000595:Amino Acid Sequence; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D003960:Diaminopimelic Acid / Q000096:biosynthesis; D050746:Dihydrodipicolinate Reductase; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D008239:Lysine / Q000096:biosynthesis; D010088:Oxidoreductases / Q000235:genetics*; D044925:Oxidoreductases Acting on CH-CH Group Donors; D012333:RNA, Messenger / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D012333:RNA, Messenger; D003960:Diaminopimelic Acid; D010088:Oxidoreductases; D050746:Dihydrodipicolinate Reductase; D044925:Oxidoreductases Acting on CH-CH Group Donors; D004262:DNA Restriction Enzymes; D001616:beta-Galactosidase; D008239:Lysine