1887

Abstract

Microcystins form a large family of small cyclic heptapeptides harbouring extensive modifications in amino acid residue composition and functional group chemistry. These peptide hepatotoxins contain a range of non-proteinogenic amino acids and unusual peptide bonds, and are typically -methylated. They are synthesized on large enzyme complexes consisting of non-ribosomal peptide synthetases and polyketide synthases in a variety of distantly related cyanobacterial genera. Here we report a 1236 bp in-frame deletion mutation in the gene of the microcystin biosynthetic pathway in nine strains of the genus . The deletion removed almost the entire -methyltransferase (NMT) domain. Strains of carrying the in-frame deletion mutation incorporated mainly dehydroalanine (Dha) into the microcystins they produce while strains with full-length genes incorporated mainly -methyldehydroalanine (Mdha). Interestingly, the strains of lacking the NMT domain also incorporated elevated amounts of -Ser, the precursor of Mdha and Dha, into the microcystin they produced relative to strains carrying functional NMT domains. We provide evidence for the in-frame deletion of the NMT domain without the co-conversion of the flanking adenylation domain. Our results demonstrate a further example of the strategies employed by cyanobacteria in the biosynthesis of microcystin variants.

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2008-04-01
2024-03-28
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References

  1. Carmichael W. W. 1994; The toxins of cyanobacteria. Sci Am 270:78–86
    [Google Scholar]
  2. Chomczynski P., Rymaszewski M. 2006; Alkaline polyethylene glycol-based method for direct PCR from bacteria, eukaryotic tissue samples, and whole blood. Biotechniques 40:454–458
    [Google Scholar]
  3. Christiansen G., Fastner J., Erhard M., Börner T., Dittmann E. 2003; Microcystin biosynthesis in Planktothrix: genes, evolution, and manipulation. J Bacteriol 185:564–572
    [Google Scholar]
  4. Dittmann E., Neilan B. A., Erhard M., von Döhren H., Börner T. 1997; Insertional mutagenesis of a peptide synthetase gene that is responsible for hepatotoxin production in the cyanobacterium Microcystis aeruginosa PCC 7806. Mol Microbiol 26:779–787
    [Google Scholar]
  5. Felsenstein J. 1993 phylip (phylogeny inference package), version 3.5. Distributed by the author. Department of Genome Sciences University of Washington; Seattle, USA:
    [Google Scholar]
  6. Fewer D. P., Rouhiainen L., Jokela J., Wahlsten M., Laakso K., Wang H., Sivonen K. 2007; Recurrent adenylation domain replacement in the microcystin synthetase gene cluster. BMC Evol Biol 7:183
    [Google Scholar]
  7. Finking R., Marahiel M. A. 2004; Biosynthesis of nonribosomal peptides. Annu Rev Microbiol 58:453–488
    [Google Scholar]
  8. Halinen K., Jokela J., Fewer D. P., Wahlsten M., Sivonen K. 2007; Direct evidence for production of microcystins by Anabaena strains from the Baltic Sea. Appl Environ Microbiol 73:6543–6550
    [Google Scholar]
  9. Harada K.-I., Ogawa K., Kimura Y., Murata H., Suzuki M., Thorn P. M., Evans W. R., Carmichael W. W. 1991; Microcystins from Anabaena flos-aquae NRC 525–17. Chem Res Toxicol 4:535–540
    [Google Scholar]
  10. Jochimsen E. M., Carmichael W. W., An J. S., Cardo D. M., Cookson S. T., Holmes C. E., Antunes M. B., de Melo Filho D. A., Lyra T. M. other authors 1998; Liver failure and death after exposure to microcystins at a hemodialysis center in Brazil. N Engl J Med 338:873–878
    [Google Scholar]
  11. Kahru M. 1997; Using satellites to monitor large-scale environmental change in the Baltic Sea. In Monitoring Algal Blooms: New Techniques for Detecting Large-Scale Environmental Change pp 43–61 Edited by Kahru M., Brown C. W. Berlin, Germany: Springer-Verlag;
    [Google Scholar]
  12. Karlsson K. M., Kankaanpää H, Huttunen M., Meriluoto J. 2005; First observation of microcystin-LR in pelagic cyanobacterial blooms in the northern Baltic Sea. Harmful Algae 4:163–166
    [Google Scholar]
  13. Kotai J. 1972 Instructions for Preparation of Modified Nutrient Solution Z8 for Algae. Norwegian Institute for Water Research Publication B-11/69 Oslo: Norwegian Institute for Water Research;
    [Google Scholar]
  14. Krishnamurthy T., Szafraniec L., Hunt D. F., Shabanowitz J., Yates J. R. III, Hauer C. R., Carmichael W. W., Skulberg O., Codd G. A., Missler S. 1989; Structural characterization of toxic cyclic peptides from blue-green algae by tandem mass spectrometry. Proc Natl Acad Sci U S A 86:770–774
    [Google Scholar]
  15. Kurmayer R., Christiansen G., Gumpenberger M., Fastner J. 2005; Genetic identification of microcystin ecotypes in toxic cyanobacteria of the genus Planktothrix. Microbiology 151:1525–1533
    [Google Scholar]
  16. Lautru S., Challis G. L. 2004; Substrate recognition by non-ribosomal peptide synthetase multi-enzymes. Microbiology 150:1629–1636
    [Google Scholar]
  17. Luukkainen R., Sivonen K., Namikoshi M., Färdig M., Rinehart K. L., Niemelä S. I. 1993; Isolation and identification of eight microcystins from thirteen Oscillatoria agardhii strains and structure of a new microcystin. Appl Environ Microbiol 59:2204–2220
    [Google Scholar]
  18. MacKintosh C., Beattie K. A., Klumpp S., Cohen P., Codd G. A. 1990; Cyanobacterial microcystin-LR is a potent and specific inhibitor of protein phosphatases 1 and 2A from both mammals and higher plants. FEBS Lett 264:187–192
    [Google Scholar]
  19. Marahiel M. A., Stachelhaus T., Mootz H. D. 1997; Modular peptide synthetases involved in nonribosomal peptide synthesis. Chem Rev 97:2651–2674
    [Google Scholar]
  20. Mikalsen B., Boison G., Skulberg O. M., Fastner J., Davies W., Gabrielsen T. M., Rudi K., Jakobsen K. S. 2003; Natural variation in the microcystin synthetase operon mcyABC and impact on microcystin production in Microcystis strains. J Bacteriol 185:2774–2785
    [Google Scholar]
  21. Moffitt M. C., Neilan B. A. 2004; Characterization of the nodularin synthetase gene cluster and proposed theory of the evolution of cyanobacterial hepatotoxins. Appl Environ Microbiol 70:6353–6362
    [Google Scholar]
  22. Namikoshi M., Sivonen K., Evans W. R., Carmichael W. W., Sun F., Rouhiainen L., Luukkainen R., Rinehart K. L. 1992a; Two new l-serine variants of microcystins-LR and -RR from Anabaena sp. strains 202 A1 and 202 A2. Toxicon 30:1457–1464
    [Google Scholar]
  23. Namikoshi M., Sivonen K., Evans W. R., Carmichael W. W., Rouhiainen L., Luukkainen R., Rinehart K. L. 1992b; Structures of three new homotyrosine-containing microcystins and a new homophenylalanine variant from Anabaena sp. strain 66. Chem Res Toxicol 5:661–666
    [Google Scholar]
  24. Namikoshi M., Yuan M., Sivonen K., Carmichael W. W., Rinehart K. L., Rouhiainen L., Sun F., Brittain S., Otsuki A. 1998; Seven new microcystins possessing two l-glutamic acid units, isolated from Anabaena sp. strain 186. Chem Res Toxicol 11:143–149
    [Google Scholar]
  25. Nishiwaki-Matsushima R., Ohta T., Nishiwaki S., Suganuma M., Kohyama K., Ishikawa T., Carmichael W. W., Fujiki H. 1992; Liver tumor promotion by the cyanobacterial cyclic peptide toxin microcystin-LR. J Cancer Res Clin Oncol 118:420–424
    [Google Scholar]
  26. Nishizawa T., Asayama M., Fujii K., Harada K., Shirai M. 1999; Genetic analysis of the peptide synthetase genes for a cyclic heptapeptide microcystin in Microcystis spp. J Biochem (Tokyo 126:520–529
    [Google Scholar]
  27. Rantala A., Fewer D. P., Hisbergues M., Rouhiainen L., Vaitomaa J., Börner T., Sivonen K. 2004; Phylogenetic evidence for the early evolution of microcystin synthesis. Proc Natl Acad Sci U S A 101:568–573
    [Google Scholar]
  28. Ressom R., Soong F. S., Fitzgerald J., Turczynowicz L., El Saadi O., Roder D., Maynard T., Falconer I. R. 1994 Health Effects of Toxic Cyanobacteria (Blue-Green Algae) Canberra: National Health and Medical Council, Australian Government Publishing Service;
  29. Robillot C., Vinh J., Puiseux-Dao S., Hennion M. C. 2000; Hepatotoxin production kinetics of the cyanobacterium Microcystis aeruginosa PCC 7820, as determined by HPLC: mass spectrometry and protein phosphatase bioassay. Environ Sci Technol 34:3372–3378
    [Google Scholar]
  30. Rouhiainen L., Vakkilainen T., Siemer B. L., Buikema W., Haselkorn R., Sivonen K. 2004; Genes coding for hepatotoxic heptapeptides (microcystins) in the cyanobacterium Anabaena strain 90. Appl Environ Microbiol 70:686–692
    [Google Scholar]
  31. Sieber S. A., Marahiel M. A. 2005; Molecular mechanisms underlying non-ribosomal peptide synthesis: approaches to new antibiotics. Chem Rev 105:715–738
    [Google Scholar]
  32. Sivonen K., Jones G. 1999; Cyanobacterial toxins. In Toxic Cyanobacteria in Water pp 41–111 Edited by Chorus I., Bartram J. London: E&FN Spon;
    [Google Scholar]
  33. Sivonen K., Namikoshi M., Evans W. R., Carmichael W. W., Sun F., Rouhiainen L., Luukkainen R., Rinehart K. L. 1992a; Isolation and characterization of a variety of microcystins from seven strains of the cyanobacterial genus Anabaena. Appl Environ Microbiol 58:2495–2500
    [Google Scholar]
  34. Sivonen K., Skulberg O. M., Namikoshi M., Evans W. R., Carmichael W. W., Rinehart K. L. 1992b; Two methyl ester derivatives of microcystins, cyclic heptapeptide hepatotoxins, isolated from Anabaena flos-aquae strain CYA 83/1. Toxicon 30:1465–1471
    [Google Scholar]
  35. Stachelhaus T., Schneider A., Marahiel M. A. 1995; Rational design of peptide antibiotics by targeted replacement of bacterial and fungal domains. Science 269:69–72
    [Google Scholar]
  36. Stal L. J., Albertano P., Bergman B., von Bröckel K., Gallon J. R., Hayes P. K., Sivonen K., Walsby A. E. 2003; BASIC: Baltic Sea cyanobacteria: an investigation of the structure and dynamics of water blooms of cyanobacteria in the Baltic Sea – responses to a changing environment. Cont Shelf Res 23:1695–1714
    [Google Scholar]
  37. Tillett D., Dittmann E., Erhard M., von Döhren H., Börner T., Neilan B. A. 2000; Structural organization of microcystin biosynthesis in Microcystis aeruginosa PCC 7806: an integrated peptide-polyketide synthetase system. Chem Biol 7:753–764
    [Google Scholar]
  38. von Döhren H., Keller U., Vater J., Zocher R. 1997; Multifunctional peptide synthetases. Chem Rev 97:2675–2705
    [Google Scholar]
  39. WHO 1998; Guidelines for Drinking Water Quality: Addendum to Volume 2, Health Criteria and Other Supporting Information. , 2nd edn. pp 1–283 Geneva: World Health Organization;
    [Google Scholar]
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