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Cloning and characterisation of genes for tetrapyrrole biosynthesis from the cyanobacterium Anacystis nidulans R2

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Abstract

The genes for 5-aminolevulinic acid dehydratase (ALAD) and uroporphyrinogen III synthase (UROS), two enzymes in the biosynthetic pathway for tetrapyrroles, were independently isolated from a plasmid-based genomic library of Anacystis nidulans R2 (also called Synechococcus sp. PCC7942), by their ability to complement Escherichia coli strains carrying mutations in the equivalent genes (hemB and hemD respectively). The identity of the genes was confirmed by comparing the appropriate enzyme activities in complemented and mutant strains. Subclones of the original plasmids that were also capable of complementing the mutants were sequenced. The inferred amino acid sequence of the cyanobacterial HemB protein indicates a significant difference in the metal cofactor requirement from the higher-plant enzymes, which was confirmed by overexpression and biochemical analysis. The organisation of the cyanobacterial hemD locus differs markedly from other prokaryotes. Two open reading frames were found immediately upstream of hemD. The product of one shows considerable similarity to published sequences from other organisms for uroporphyrinogen III methylase (UROM), an enzyme involved in the production of sirohaem and cobalamins (including vitamin B-12). The product of the other shows motifs which are similar to those found in proteins responsible for metabolic regulation in yeast and indicates that this family of transcription control proteins, which has previously been reported only from eukaryotes, is also represented in prokaryotes.

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References

  1. Alefounder PR, Abel C, Battersby AR: The sequence of hemC hemD and two additional E. coli genes: Nucl Acids Res 16: 9871 (1988).

    PubMed  Google Scholar 

  2. Allen MM, Stanier RY: Selective isolation of blue green algae from water and soil. J Gen Microbiol 51: 203–209 (1968).

    PubMed  Google Scholar 

  3. Blanche F, Robin C, Couder M, Faucher D, Cauchois L, Cameron B, Crouzet J: Purification, characterisation and molecular cloning of S-adenosyl-L-methionine Uroporphyrinogen III methyltransferase from Methanobacterium ivanovii. J Bact 173: 4637–4645 (1991).

    PubMed  Google Scholar 

  4. Boese QF, Spano AJ, Li JM, Timko MP: Aminolevulinic acid dehydratase in pea (Pisum sativum L) identification of an unusual metal binding domain in the plant enzyme. J Biol Chem 226: 17060–17066 (1991).

    Google Scholar 

  5. Chartrand P, Tardiff D, Sasarman A: Uroporphyrin-and coproporphyrin I accumulating mutant of Escherichia coli K12. J Gen Microbiol 110: 61–66 (1979).

    PubMed  Google Scholar 

  6. Crozet, Cauchois L, Blanche F, Debussche L, Thibaut D, Rouyez M-C, Rigault S, Mayaux J-F, Cameron B: Nucleotide sequence of Pseudomonas denitrificans 5.4 kilobase DNA fragment containing five cob genes and identification of structural genes encoding S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase and cobyrinic acid a,c-diamide synthase. J Bact 172: 5968–5979 (1990).

    PubMed  Google Scholar 

  7. Deveraux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).

    PubMed  Google Scholar 

  8. Dunn MF, Pattison SE, Storm MC, Quiel E: Comparison of zinc binding domains in the 7S nerve growth factor and the zinc-insulin hexamer. Biochemistry 19: 718–725 (1980).

    PubMed  Google Scholar 

  9. Echelard Y, Dymetryszyn J, Drolet M, Sasarman A: Nucleotide sequence of the hemB gene of Escherichia coli K12. Mol Gen Genet 214: 503–508 (1988).

    Article  PubMed  Google Scholar 

  10. Fisher R, Tuli R, Haselkorn R: A cloned cyanobacterial gene for glutamine synthetase functions in Escherichia coli, but the enzyme is not adenylylated. Proc Natl Acad Sci USA 78: 3393–3397 (1981).

    PubMed  Google Scholar 

  11. Geoghegan CM, Houghton JA: Molecular cloning of a cyanobacterial gene which increases the UV and methyl methanesulphonate survival of recA strains of Escherichia coli K12. J Gen Microbiol 133: 119–126 (1987).

    PubMed  Google Scholar 

  12. Gibbs PNB, Chaudhry AG, Jordan PM: Purification and properties of 5-aminolaevulinate dehydratase from human erythrocytes. Biochem J 230: 25–34 (1985).

    PubMed  Google Scholar 

  13. Gibbs PNB, Jordan PM: Identification of lysine at the active site of human 5-aminolaevulinate dehydratase. Biochem J 236: 447–451 (1986).

    PubMed  Google Scholar 

  14. Gibbs PNB, Jordan PM: 5-aminolaevulinate dehydratase: zinc-65 binding and exchange with the enzyme from human erythrocytes. Biochem Soc Trans 9: 232–233 (1981).

    Google Scholar 

  15. Grimm B, Bull A, Breu V: Structural genes of glutamate 1 semialdehyde aminotransferase for porphyrin synthesis in a cyanobacterium and Escherichia coli. Mol Gen Genet 225: 1–10 (1991).

    Article  PubMed  Google Scholar 

  16. Hansson M, Rutberg L, Schroder I, Hederstedt L: The Bacillus subtilis hemAXCDBL gene cluster, which encodes enzymes of the biosynthetic pathway from glutamate to Uroporphyrinogen III. J Bact 173: 2590–2599 (1991).

    PubMed  Google Scholar 

  17. Hart GJ, Battersby AR: Purification and properties of uroporphyrinogen III synthase (co-synthase) from Euglena gracilis. Biochem J 232: 151–160 (1985).

    PubMed  Google Scholar 

  18. Jordan PM, Nordlov H, Burton G, Scott AI: Nucleotide sequence for the hemD gene of Escherichia coli encoding Uroporphyrinogen III synthase and initial evidence for a hem operon. FEBS Lett 115: 269–272 (1980).

    Article  PubMed  Google Scholar 

  19. Kiel JAKW, Boels JM, Beldman G, Venema G: Nucleotide sequence of the Synechococcus sp. PCC7942 branching enzyme gene (glgB): expression in Bacillus subtilis. Gene 89: 77–84 (1990).

    Article  PubMed  Google Scholar 

  20. Kodaki T, Katagari F, Asano M, Izui K, Katsuki H: Cloning of phosphoenolpyruvate carboxylase (EC 4.1.1.31) gene from a cyanobacterium. J Biochem (Tokyo) 97: 533–540 (1985).

    Google Scholar 

  21. Koenen M: Recovery of DNA from agarose gels using liquid nitrogen. Trends Genet 5: 137 (1989).

    Google Scholar 

  22. Legrain P, Chapon C, Galisson F: Proteins involved in mitosis, RNA synthesis and premRNA splicing share a common repeating motif. Nucl Acids Res 19: 2509–2510 (1991).

    PubMed  Google Scholar 

  23. Liedgens W, Lutz C, Schneider HAW: Molecular properties of 5-aminolevulinic acid dehydratase from Spinacia oleracia. Eur J Biochem 135: 75–79 (1983).

    PubMed  Google Scholar 

  24. Li J-M, Umanoff H, Proenca R, Russell CS, Cosloy SD: Cloning of the Escherichia coli K-12 hemB gene. J Bact 170: 1021–1025 (1988).

    PubMed  Google Scholar 

  25. Lim CJ, Gleason FK, Fuchs JA: Cloning, expression and characterisation of the Anabaena thioredoxin gene in Escherichia coli. J. Bact 168: 1258–1264 (1986).

    PubMed  Google Scholar 

  26. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  27. Manzano C, Candau P, Gomez-Moreno C, Relimpio AM, Losada M: Ferredoxin dependent photosynthetic reduction of nitrate and nitrite by particles of Anacystis nidulans. Mol Cell Biochem 10: 161–169 (1976).

    PubMed  Google Scholar 

  28. Marmur J: A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3: 208–218 (1961).

    Google Scholar 

  29. Messing J, Bankier AT: The use of single-stranded DNA phage in DNA sequencing. In: Howe CJ, Ward ES (eds) Nucleic Acids Sequencing: A Practical Approach, pp. 1–36. IRL Press, Oxford, UK (1989).

    Google Scholar 

  30. Murphy RC, Bryant DA, Porter RD, Tandeau de Marsac N: Molecular cloning and characterisation of the rec A gene from the cyanobacterium Synechococcus sp. strain PCC7002. J Bact 169: 2739–2747 (1987).

    PubMed  Google Scholar 

  31. Murphy RC, Gasparich GE, Bryant DA, Porter RD: Nucleotide sequence and further characterisation of the Synechococcus sp. strain PCC7002 rec A gene: complementation of a cyanobacterial rec A mutation by the Escherichia c coli recA gene. J. Bact 172: 967–976 (1990).

    PubMed  Google Scholar 

  32. Pearson WR, Lipman DJ: Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85: 2444–2448 (1988).

    PubMed  Google Scholar 

  33. Robin C, Blanche F, Cauchois L, Cameron D, Couder M, Crouzet J: Primary structure, expression in Escherichia coli and properties of S-adenosyl-L-methionine Uroporphyrinogen III methyltransferase from Bacillus megaterium. J Bact 173: 4893–4896 (1991).

    PubMed  Google Scholar 

  34. Sasarman A, Desroches M: Uroporphyrinogen III cosynthase-deficient mutant of Salmonella typhimurium LT2. J Bact 128: 717–721 (1976).

    PubMed  Google Scholar 

  35. Sasarman A, Horodniceau T: Locus determining normal colony formation on the chromosome of Escherichia coli K12. J Bact 94: 1268–1269 (1967).

    PubMed  Google Scholar 

  36. Sasarman A, Sanderson KE, Surdeanu M, Sonea S: Hemin deficient mutants of Salmonella typhimurium. J Bact 102: 531–536 (1970).

    PubMed  Google Scholar 

  37. Schaumberg A, Schneider-Poetsch HAW, Eckerskorn C: Characterisation of plastid 5-aminolevulinate dehydratase (ALAD EC 4.21.24) from spinach (Spinacia oleracia L) by sequencing and comparison with non-plant enzymes. Z Naturforsch C 47: 77–84 (1992).

    PubMed  Google Scholar 

  38. Schneider HAW: Anreicherung und Eigenschaften von ∂-Aminolävulinate-dehydratase aus Spinat (Spinacia oleracia). Z Pflanzenphysiol 62: 328–342 (1970).

    Google Scholar 

  39. Schultz J, Marshal-Carlson L, Carlson M: The N-terminal TPR region is the functional domain of SSN6 a nuclear phosphoprotein of Saccharomyces cerevisiae. Mol Cell Biol 10: 4744–4756 (1990).

    PubMed  Google Scholar 

  40. Sikorski RS, Boguski MS, Goebl M, Hieter P: A repeating amino acid motif in CDC23 defines a family of proteins and a new relationship among genes required for mitosis and RNA synthesis. Cell 60: 307–317 (1990).

    PubMed  Google Scholar 

  41. Smith AG: Subcellular localisation of two porphyrin synthesis genes in Pisum sativum (pea) and Arum (cuckoopint) species. Biochem J 249: 423–428 (1988).

    PubMed  Google Scholar 

  42. Spencer P, Jordan PM: Purification and characterization of a 5-aminolaevulinic acid dehydratase from Escherichia coli and a study of the reactive thiols at the metal-binding domain. Biochem J 290: 279–287 (1993).

    PubMed  Google Scholar 

  43. Tabor S, Richardson CC: Selective inactivation of the exonuclease activity of bacteriophage T7 DNA polymerase by in vitro mutagenesis. J Biol Chem 264: 6447–6458 (1989).

    PubMed  Google Scholar 

  44. Tomioka N, Sugiura M: The complete nucleotide sequence of a 16S ribosomal RNA gene from a blue-green alga, Anacystis nidulans. Mol Gen Genet 191: 46–50 (1983).

    PubMed  Google Scholar 

  45. Trumbly RJ: Cloning and characterisation of the CYC8 gene mediating glucose repression in yeast. Gene 73: 97–111 (1988).

    PubMed  Google Scholar 

  46. Warren MJ, Scott AI: Tetrapyrrole assembly and modification into the ligands of biologically active cofactors. Trends Biochem Sci 15: 486–491 (1990).

    PubMed  Google Scholar 

  47. Wilson EL, Burger PE, Dowdle EB: Beef-liver 5-aminolevulinic acid dehydratase purification and properties. Eur J Biochem 29: 563–571 (1972).

    PubMed  Google Scholar 

  48. Yabuki Y, Mori E, Tamura G: Nitrite reductase in the cyanobacterium Spirulina platensis. Agric Biol Chem 49: 3061–3062 (1985).

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, University of Cambridge, Tennis Court Road, CB2 1QW, Cambridge, UK

    Matthew C. Jones & Christopher J. Howe

  2. Department of Plant Sciences, University of Cambridge, Downing Street, CB2 3EA, Cambridge, UK

    Joanne M. Jenkins & Alison G. Smith

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  1. Matthew C. Jones
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  3. Alison G. Smith
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  4. Christopher J. Howe
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Jones, M.C., Jenkins, J.M., Smith, A.G. et al. Cloning and characterisation of genes for tetrapyrrole biosynthesis from the cyanobacterium Anacystis nidulans R2. Plant Mol Biol 24, 435–448 (1994). https://doi.org/10.1007/BF00024112

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  • DOI: https://doi.org/10.1007/BF00024112

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