Characterization of dapB, a gene required by Pseudomonas syringae pv. tabaci BR2.024 for lysine and tabtoxinine-beta-lactam biosynthesis.
The dapB gene, which encodes L-2,3-dihydrodipicolinate reductase, the second enzyme of the lysine branch of the aspartic amino acid family, was cloned and sequenced from a tabtoxin-producing bacterium, Pseudomonas syringae pv. tabaci BR2.024. The deduced amino acid sequence shared 60 to 90% identity to known dapB gene products from gram-negative bacteria and 19 to 21% identity to the dapB products from gram-positive bacteria. The consensus sequence for the NAD(P)H binding site [(V/I)(A/G)(V/I)XGXXGXXG)] and the proposed substrate binding site (HHRHK) were conserved in the polypeptide. A BR2.024 dapB mutant is a diaminopimelate auxotroph and tabtoxin negative. The addition of a mixture of L-,L-, D,D-, and meso-diaminopimelate to defined media restored growth but not tabtoxin production. Cloned DNA fragments containing the parental dapB gene restored the ability to grow in defined media and tabtoxin production to the dapB mutant. These results indicate that the dapB gene is required for both lysine and tabtoxin biosynthesis, thus providing the first genetic evidence that the biosynthesis of tabtoxin proceeds in part along the lysine biosynthetic pathway. These data also suggest that L-2,3,4,5-tetrahydrodipicolinate is a common intermediate for both lysine and tabtoxin biosynthesis.
Liu
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Department of Crop Sciences, University of Illinois at Urbana-Champaign, 61801, USA.
Shaw
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PD
D000595:Amino Acid Sequence; D001384:Azetidines / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000737:chemistry; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases; D004269:DNA, Bacterial; D050746:Dihydrodipicolinate Reductase; D004151:Dipeptides / Q000096:biosynthesis; D017353:Gene Deletion; D005816:Genetic Complementation Test; D008239:Lysine / Q000096:biosynthesis* / Q000737:chemistry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016254:Mutagenesis, Insertional; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D019895:Transposases
D001384:Azetidines; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004151:Dipeptides; C051368:tabtoxinine beta-lactam; D010088:Oxidoreductases; D050746:Dihydrodipicolinate Reductase; D044925:Oxidoreductases Acting on CH-CH Group Donors; D004254:DNA Nucleotidyltransferases; D019895:Transposases; C003854:tabtoxin; D008239:Lysine