Summary
The gene coding for the phaseolotoxin-insensitive ornithine carbamoyltransferase (OCTase) fromPseudomonas syringae pv.phaseolicola has been cloned and sequenced. The gene has a deduced coding capacity for a polypeptide with a calculated M, of 36520 daltons. Comparison of the amino acid sequence of the OCTase enzymes encoded by theP. aeruginosa argF and theEscherichia coli argI andargF genes with the deduced sequence of the newly identified gene shows that 79 amino acid residues are strictly conserved in all four polypeptides; among these 7 out of 9 residues are involved in enzyme function. Of three amino acid regions that have been implicated in substrate binding or catalysis, two are strictly conserved, and the third involved in carbamoylphosphate binding differs. This correlates well with published data showing that phaseolotoxin competes for the carbamoylphosphate binding site in the phaseolotoxin-sensitive OCTases. We propose that the gene be namedargK.
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References
Bencini DA, Houghton JE, Hoover TA, Foltermann KF, Wild JR, O'Donovan GA (1983) The DNA sequence ofargI fromEscherichia coli K12. Nucleic Acids Res 11:8509–8518
Casadaban MJ, Martinez-Arias A, Shapira SK, Chou J (1983) β-Galactosidase gene fusions for analyzing gene expression inEscherichia coli and yeast. Methods Enzymol 100:293–308
Ceriotti G (1974) ornithine carbamoyltransferase. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol 2. Verlag Chemie International, Deerfield Beach, Florida, pp 691–698
Cunin R, Eckhardt T, Piette J, Boyen A, Pierard A, Glansdorff N (1983) Molecular basis for modulated regulation of gene expression in the arginine regulon ofEscherichia coli K-12. Nucleic Acids Res 11:5007–5019
Durbin RD, Langston-Unkefer PJ (1988) The mechanisms for self-protection against bacterial phytotoxins. Annu Rev Phytopathol 26:313–329
Ferguson AR, Johnston JS, Mitchell RE (1980) Resistance ofPseudomonas syringae pv.phaseolicola to its own toxin, phaseolotoxin. FEMS Microbiol Lett 7:123–125
Frantz B, Chakrabarty AM (1986) Degradative plasmids inPseudomonas. In: Sokatch JR (ed) The bacteria, vol X. Academic Press, NY, pp 295–323
Friedman DI, Imperiale MJ, Adhya S (1987) RNA 3′ end formation in the control of gene expression. Annu Rev Genet 21:453–488
Hawley DK, McClure WR (1983) Compilation and analysis ofEscherichia coli promoter DNA sequences. Nucleic Acids Res 11:2237–2255
Itoh Y, Soldati L, Stalon V, Falmagne P, Terawaki Y, Leisinger T, Haas D (1988) Anabolic ornithine carbamoyltransferase ofPseudomonas aeruginosa: nucleotide sequence and transcriptional control of theargF structural gene. J Bacteriol 170:2725–2734
Lelliott RA, Stead DE (1987) Methods for the diagnosis of bacterial diseases of plants. In: Preece TF (ed) Methods in plant pathology, vol 2. Blackwell Scientific Publications, Oxford, pp 176–177
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78
Minton NP (1984) Improved plasmid vectors for the isolation of translationallacZ gene fusions. Gene 31:269–273
Mitchell RE (1984) The relevance of non-host-specific toxins in the expression of virulence by pathogens. Annu Rev Phytopathol 22:215–245
Panopoulos NJ, Peet RC (1985) The molecular genetics of plant pathogenic bacteria and their plasmids. Annu Rev Phytopathol 23:381–419
Peet RC, Panopoulos NJ (1987) Ornithine carbamoyltransferase genes and phaseolotoxin immunity inPseudomonas syringae pv.phaseolicola. EMBO J 6:3585–3591
Peet RC, Lindgren PB, Willis DK, Panopoulos NJ (1986) Identification and cloning of genes involved in phaseolotoxin production bypseudomonas syringae pv.phaseolicola. J Bacteriol 166:1096–1105
Rozkot F, Sazelova P, Pivec L (1989) A novel method for promoter search enhanced by function-specific subgrouping of promoters developed and tested onE. coli system. Nucleic Acids Res 17:4799–4815
Staskawicz BJ, Panopoulos NJ (1979) A rapid and sensitive microbiological assay for phaseolotoxin. Phytopathology 69:663–666
Staskawicz BJ, Panopoulos NJ, Hoogenraad NJ (1980) Phaseolotoxin-insensitive ornithine carbamoyltransferase ofPseudomonas syringae pv.phaseolicola: Basis for immunity to phaseolotoxin. J Bacteriol 142:720–723
Stebbins JW, Xu W, Kantrowitz ER (1989) Three residues involved in binding and catalysis in the carbamyl phosphate binding site ofEscherichia coli aspartate transcarbamylase. Biochemistry 28:2592–2600
Stormo GD, Schneider TD, Gold LM (1982) Characterization of translational initiation sites inE. coli. Nucleic Acids Res 10:2971–2996
Templeton MD, Sullivan PA, Shepherd MG (1986) Phaseolotoxin-insensitivel-ornithine transcarbamoylase fromPseudomonas syringae pv.phaseolicola. Physiol Mol Plant Pathol 29:393–403
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
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Mosqueda, G., Van den Broeck, G., Saucedo, O. et al. Isolation and characterization of the gene fromPseudomonas syringae pv.phaseolicola encoding the phaseolotoxin-insensitive ornithine carbamoyltransferase. Mol Gen Genet 222, 461–466 (1990). https://doi.org/10.1007/BF00633857
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DOI: https://doi.org/10.1007/BF00633857