A Possible Role for Acetylated Intermediates in Diaminopimelate and Tabtoxinine-beta-lactam Biosynthesis in Pseudomonas Syringae Pv. Tabaci BR2.024

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1997 Sep 19

PMID 9294453

Citations 1

Authors

L Liu

P D SHAW

Affiliations

Soon will be listed here.

Abstract

The deduced product of an open reading frame (ORF3) located in the tabtoxinine-beta-lactam (T beta L) biosynthetic region of Pseudomonas syringae pv. tabaci BR2.024 (BR2.024) has significant sequence homology to the dapD products of other bacteria. dapD encodes L-2,3,4,5-tetrahydrodipicolinate succinyl coenzyme A succinyltransferase (THDPA-ST), an enzyme in the diaminopimelate (DAP) and lysine biosynthetic pathway. Complementation studies, in vitro transcription-translation experiments, and enzymatic assays indicated that ORF3 encodes a product with THDPA-ST activity in Escherichia coli dapD mutant beta 274. However, a BR2.024 mutant with an insert in ORF3 was prototrophic, and only basal THDPA-ST activity was detected in extracts of both parent and mutant. This finding suggested that ORF3 was not required for DAP biosynthesis and that it did not encode a product with THDPA-ST activity. The results of enzymatic studies, indicating that BR2.024 uses acetylated intermediates for DAP biosynthesis, are consistent with the hypothesis that BR2.024 does not need THDPA-ST for DAP biosynthesis. The ORF3 mutant produced reduced levels of tabtoxin, indicating that ORF3 may have a role in T beta L biosynthesis. We have named the gene tabB and have proposed a possible function for the gene product.

Citing Articles

Pseudomonas syringae phytotoxins: mode of action, regulation, and biosynthesis by peptide and polyketide synthetases.

Bender C, Gross D Microbiol Mol Biol Rev. 1999; 63(2):266-92.

PMID: 10357851 PMC: 98966. DOI: 10.1128/MMBR.63.2.266-292.1999.

References

WOOLLEY D, Pringle R, Braun A . Isolation of the phytopathogenic toxin of Pseudomonas tabaci, an antagonist of methionine. J Biol Chem. 1952; 197(1):409-17. View

Martin C, Cami B, Yeh P, Stragier P, Parsot C, PATTE J . Pseudomonas aeruginosa diaminopimelate decarboxylase: evolutionary relationship with other amino acid decarboxylases. Mol Biol Evol. 1988; 5(5):549-59. DOI: 10.1093/oxfordjournals.molbev.a040515. View

Cassan M, Parsot C, COHEN G, PATTE J . Nucleotide sequence of lysC gene encoding the lysine-sensitive aspartokinase III of Escherichia coli K12. Evolutionary pathway leading to three isofunctional enzymes. J Biol Chem. 1986; 261(3):1052-7. View

Vogel H, Bonner D . Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956; 218(1):97-106. View

Ishino S, Mizukami T, Yamaguchi K, Katsumata R, Araki K . Nucleotide sequence of the meso-diaminopimelate D-dehydrogenase gene from Corynebacterium glutamicum. Nucleic Acids Res. 1987; 15(9):3917. PMC: 340791. DOI: 10.1093/nar/15.9.3917. View

Piwowarski J, SHAW P . Characterization of plasmids from plant pathogenic pseudomonads. Plasmid. 1982; 7(1):85-94. DOI: 10.1016/0147-619x(82)90030-0. View

Taylor P, Schnoes H, Durbin R . Characterization of chlorosis-inducing toxins from a plant pathogenic Pseudomonas Sp. Biochim Biophys Acta. 1972; 286(1):107-17. DOI: 10.1016/0304-4165(72)90096-7. View

Bouvier J, Richaud C, Higgins W, Bogler O, Stragier P . Cloning, characterization, and expression of the dapE gene of Escherichia coli. J Bacteriol. 1992; 174(16):5265-71. PMC: 206361. DOI: 10.1128/jb.174.16.5265-5271.1992. View

Liu L, SHAW P . Characterization of dapB, a gene required by Pseudomonas syringae pv. tabaci BR2.024 for lysine and tabtoxinine-beta-lactam biosynthesis. J Bacteriol. 1997; 179(2):507-13. PMC: 178722. DOI: 10.1128/jb.179.2.507-513.1997. View

10.

Richaud C, Printz C . Nucleotide sequence of the dapF gene and flanking regions from Escherichia coli K12. Nucleic Acids Res. 1988; 16(21):10367. PMC: 338869. DOI: 10.1093/nar/16.21.10367. View

11.

Yeh P, Sicard A, Sinskey A . General organization of the genes specifically involved in the diaminopimelate-lysine biosynthetic pathway of Corynebacterium glutamicum. Mol Gen Genet. 1988; 212(1):105-11. DOI: 10.1007/BF00322451. View

12.

Thomas M, Langston-Unkefer P, Uchytil T, Durbin R . Inhibition of Glutamine Synthetase from Pea by Tabtoxinine-beta-lactam. Plant Physiol. 1983; 71(4):912-5. PMC: 1066143. DOI: 10.1104/pp.71.4.912. View

13.

Richaud F, Richaud C, Ratet P, PATTE J . Chromosomal location and nucleotide sequence of the Escherichia coli dapA gene. J Bacteriol. 1986; 166(1):297-300. PMC: 214591. DOI: 10.1128/jb.166.1.297-300.1986. View

14.

Fellay R, Krisch H, Prentki P, Frey J . Omegon-Km: a transposable element designed for in vivo insertional mutagenesis and cloning of genes in gram-negative bacteria. Gene. 1989; 76(2):215-26. DOI: 10.1016/0378-1119(89)90162-5. View

15.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

16.

Keen N, Tamaki S, Kobayashi D, Trollinger D . Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988; 70(1):191-7. DOI: 10.1016/0378-1119(88)90117-5. View

17.

Kinscherf T, Coleman R, Barta T, Willis D . Cloning and expression of the tabtoxin biosynthetic region from Pseudomonas syringae. J Bacteriol. 1991; 173(13):4124-32. PMC: 208062. DOI: 10.1128/jb.173.13.4124-4132.1991. View

18.

Riddles P, Blakeley R, ZERNER B . Ellman's reagent: 5,5'-dithiobis(2-nitrobenzoic acid)--a reexamination. Anal Biochem. 1979; 94(1):75-81. DOI: 10.1016/0003-2697(79)90792-9. View

19.

Vuorio R, Harkonen T, Tolvanen M, Vaara M . The novel hexapeptide motif found in the acyltransferases LpxA and LpxD of lipid A biosynthesis is conserved in various bacteria. FEBS Lett. 1994; 337(3):289-92. DOI: 10.1016/0014-5793(94)80211-4. View

20.

Figurski D, Helinski D . Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979; 76(4):1648-52. PMC: 383447. DOI: 10.1073/pnas.76.4.1648. View