1887

Abstract

A unique -to- racemization of arginine by coupled arginine dehydrogenases DauA and DauB encoded by the operon has been recently reported as a prerequisite for -arginine utilization as the sole source of carbon and nitrogen through -arginine catabolic pathways in . In this study, enzymic properties of the catabolic FAD-dependent -amino acid dehydrogenase DauA and the physiological functions of the operon were further characterized with other -amino acids. These results establish DauA as a -amino acid dehydrogenase of broad substrate specificity, with -Arg and -Lys as the two most effective substrates, based on the kinetic parameters. In addition, expression of is specifically induced by exogenous -Arg and -Lys, and mutations in the operon affect utilization of these two amino acids alone. The function of DauR as a repressor in the control of the operon was demonstrated by promoter activity measurements and mobility shift assays with purified His-tagged protein . The potential effect of 2-ketoarginine (2-KA) derived from -Arg deamination by DauA as a signal molecule in induction was first revealed by mutation analysis and further supported by its effect on alleviation of DauR–DNA interactions. Through sequence analysis, putative DauR operators were identified and confirmed by mutation analysis. Induction of the operon to the maximal level was found to require the -arginine-responsive regulator ArgR, as supported by the loss of inductive effect by -Arg on expression in the mutant and binding of purified ArgR to the regulatory region . In summary, this study establishes that optimal induction of the operon requires relief of DauR repression by 2-KA and activation of ArgR by -Arg as a result of -Arg racemization by the encoded DauA and DauB.

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2010-01-01
2024-12-14
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References

  1. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  2. Caparros M., Pisabarro A. G., de Pedro M. A. 1992; Effect of d-amino acids on structure and synthesis of peptidoglycan in Escherichia coli. J Bacteriol 174:5549–5559
    [Google Scholar]
  3. Challis G. L., Naismith J. H. 2004; Structural aspects of non-ribosomal peptide biosynthesis. Curr Opin Struct Biol 14:748–756
    [Google Scholar]
  4. Erikson O., Hertzberg M., Nasholm T. 2004; A conditional marker gene allowing both positive and negative selection in plants. Nat Biotechnol 22:455–458
    [Google Scholar]
  5. Farinha M. A., Kropinski A. M. 1990; Construction of broad-host-range plasmid vectors for easy visible selection and analysis of promoters. J Bacteriol 172:3496–3499
    [Google Scholar]
  6. Friede J. D., Henderson L. M. 1976; Metabolism of 5-hydroxylysine in Pseudomonas fluorescens. J Bacteriol 127:1239–1247
    [Google Scholar]
  7. Gallegos M. T., Schleif R., Bairoch A., Hofmann K., Ramos J. L. 1997; Arac/XylS family of transcriptional regulators. Microbiol Mol Biol Rev 61:393–410
    [Google Scholar]
  8. Haas D., Holloway B. W., Schambock A., Leisinger T. 1977; The genetic organization of arginine biosynthesis in Pseudomonas aeruginosa. Mol Gen Genet 154:7–22
    [Google Scholar]
  9. Jann A., Stalon V., Wauven C. V., Leisinger T., Haas D. 1986; N2-Succinylated intermediates in an arginine catabolic pathway of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A 83:4937–4941
    [Google Scholar]
  10. Jann A., Matsumoto H., Haas D. 1988; The fourth arginine catabolic pathway of Pseudomonas aeruginosa. J Gen Microbiol 134:1043–1053
    [Google Scholar]
  11. Kamio M., Ko K. C., Zheng S., Wang B., Collins S. L., Gadda G., Tai P. C., Derby C. D. 2009; The chemistry of escapin: identification and quantification of the components in the complex mixture generated by an l-amino acid oxidase in the defensive secretion of the sea snail Aplysia californica. Chemistry 15:1597–1603
    [Google Scholar]
  12. Koshland D. E. 2002; The application and usefulness of the ratio kcat/ KM . Bioorg Chem 30:211–213
    [Google Scholar]
  13. Li C., Lu C. D. 2009; Arginine racemization by coupled catabolic and anabolic dehydrogenases. Proc Natl Acad Sci U S A 106:906–911
    [Google Scholar]
  14. Lu C. D. 2006; Pathways and regulation of bacterial arginine metabolism and perspectives for obtaining arginine overproducing strains. Appl Microbiol Biotechnol 70:261–272
    [Google Scholar]
  15. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  16. Nakada Y., Itoh Y. 2002; Characterization and regulation of the gbuA gene, encoding guanidinobutyrase in the arginine dehydrogenase pathway of Pseudomonas aeruginosa PAO1. J Bacteriol 184:3377–3384
    [Google Scholar]
  17. Nakada Y., Itoh Y. 2003; Identification of the putrescine biosynthetic genes in Pseudomonas aeruginosa and characterization of agmatine deiminase and N-carbamoylputrescine amidohydrolase of the arginine decarboxylase pathway. Microbiology 149:707–714
    [Google Scholar]
  18. Nishijyo T., Park S. M., Lu C. D., Itoh Y., Abdelal A. T. 1998; Molecular characterization and regulation of an operon encoding a system for transport of arginine and ornithine and the ArgR regulatory protein in Pseudomonas aeruginosa. J Bacteriol 180:5559–5566
    [Google Scholar]
  19. Park S. M., Lu C. D., Abdelal A. T. 1997; Cloning and characterization of argR, a gene that participates in regulation of arginine biosynthesis and catabolism in Pseudomonas aeruginosa PAO1. J Bacteriol 179:5300–5308
    [Google Scholar]
  20. Revelles O., Wittich R. M., Ramos J. L. 2007; Identification of the initial steps in d-lysine catabolism in Pseudomonas putida. J Bacteriol 189:2787–2792
    [Google Scholar]
  21. Schell M. J., Molliver M. E., Snyder S. H. 1995; d-Serine, an endogenous synaptic modulator: localization to astrocytes and glutamate-stimulated release. Proc Natl Acad Sci U S A 92:3948–3952
    [Google Scholar]
  22. Shin I., Wachtel E., Roth E., Bon C., Silman I., Weiner L. 2002; Thermal denaturation of Bungarus fasciatus acetylcholinesterase: is aggregation a driving force in protein unfolding?. Protein Sci 11:2022–2032
    [Google Scholar]
  23. Takahashi E., Furui M., Seko H., Shibatani T. 1997; d-Lysine production from l-lysine by successive chemical racemization and microbial asymmetric degradation. Appl Microbiol Biotechnol 47:347–351
    [Google Scholar]
  24. Takahashi K., Uchida C., Shin R. W., Shimazaki K., Uchida T. 2008; Prolyl isomerase, Pin1: new findings of post-translational modifications and physiological substrates in cancer, asthma and Alzheimer's disease. Cell Mol Life Sci 65:359–375
    [Google Scholar]
  25. Vander Wauven C., Pierard A., Kley-Raymann M., Haas D. 1984; Pseudomonas aeruginosa mutants affected in anaerobic growth on arginine: evidence for a four-gene cluster encoding the arginine deiminase pathway. J Bacteriol 160:928–934
    [Google Scholar]
  26. Vlessis A. A., Bartos D., Trunkey D. 1990; Importance of spontaneous α-ketoacid decarboxylation in experiments involving peroxide. Biochem Biophys Res Commun 170:1281–1287
    [Google Scholar]
  27. Vollmer W., Blanot D., de Pedro M. A. 2008; Peptidoglycan structure and architecture. FEMS Microbiol Rev 32:149–167
    [Google Scholar]
  28. Yang Z., Lu C. D. 2007; Functional genomics enables identification of genes of the arginine transaminase pathway in Pseudomonas aeruginosa. J Bacteriol 189:3945–3953
    [Google Scholar]
  29. Yang W., Steitz T. A. 1995; Crystal structure of the site-specific recombinase γδ resolvase complexed with a 34 bp cleavage site. Cell 82:193–207
    [Google Scholar]
  30. Yoshimura T., Esak N. 2003; Amino acid racemases: functions and mechanisms. J Biosci Bioeng 96:103–109
    [Google Scholar]
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