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Cloning and characterization of the chlorophyll biosynthesis gene chlM from Synechocystis PCC 6803 by complementation of a bacteriochlorophyll biosynthesis mutant of Rhodobacter capsulatus

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Abstract

A bacteriochlorophyll a biosynthesis mutant of the purple photosynthetic bacterium Rhodobacter capsulatus was functionally complemented with a cosmid genomic library from Synechocystis sp. PCC 6803. The complemented R. capsulatus strain contains a defined mutation in the bchM gene that codes for Mg-protoporphyrin IX methyltransferase, the enzyme which converts Mg-protoporphyrin IX to Mg-protoporphyrin IX methylester using S-adenosyl-l-methionine as a cofactor. Since chlorophyll biosynthesis also requires the same methylation reaction, the Synechocystis genome should similarly code for a Mg-protoporphyrin IX methyltransferase. Sequence analysis of the complementing Synechocystis cosmid indicates that it contains an open reading frame exhibiting 29% sequence identity to BchM. In addition, expression of the Synechocystis gene in the R. capsulatus bchM mutant via the strong R. capsulatus puc promoter was shown to support nearly wild-type levels of bacteriochlorophyll a synthesis. To our knowledge, the Synechocystis sequence thus represents the first chlorophyll biosynthesis gene homolog of bchM. The complementing Synechocystis cosmid was also shown to code for a gene product that is a member of a highly conserved family of RNA binding proteins, the function of which in cyanobacteria remains undetermined.

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References

  1. Alberti M, Burke DE, Hearst JE: Structure and sequence of the photosynthesis gene cluster. In: Blankenship RE, Madigan M, Bauer CE (eds) Anoxygenic Photosynthetic Bacteria, pp. 1082–1106. Kluwer Academic Publishers, Dordrecht, The Netherlands (1995).

    Google Scholar 

  2. Bauer CE, Bollivar DW, Suzuki JY: Genetic analysis of photopigment biosynthesis in eubacteria: a guiding light for algae and plants. J Bact 175: 3919–3925 (1993).

    PubMed  Google Scholar 

  3. Bauer CE, Buggy JJ, Mosley C: Control of photosystem genes in Rhodobacter capsulatus. Trends Genet 9: 56–60 (1993).

    Article  PubMed  Google Scholar 

  4. Bauer CE: Regulatory circuits controlling photosystem gene expression. In: Blankenship RE, Madigan M, Bauer CE (eds) Anoxygenic Photosynthetic Bacteria, pp. 1221–1234. Kluwer Academic Publishers, Dordrecht, The Netherlands (1995).

    Google Scholar 

  5. Biel AJ, Marrs BL: Transcriptional regulation of several genes for bacteriochlorophyll synthesis in Rhodopseudomonas capsulata in response to oxygen. J Bact 156: 686–694 (1983).

    PubMed  Google Scholar 

  6. Bollivar DW, Wang S, Allen JP, Bauer CE: Molecular genetic analysis of terminal steps in bacteriochlorophyll a biosynthesis: characterization of a Rhodobacter capsulatus strain that synthesizes geranylgeranyl esterified bacteriochlorophyll a. Biochemistry 33: 12763–12768 (1993).

    Google Scholar 

  7. Bollivar DW, Beale SI: Formation of the isocyclic ring of chlorophyll by isolated Chlamydomonas reinhardtii chloroplasts. Photosynth Res 43: 113–124 (1995).

    Google Scholar 

  8. Bolivar DW, Jiang ZY, Bauer CE, Beale SI: Heterologous overexpression of the bchM gene product from Rhodobacter capsulatus and demonstration that it encodes S-adenosyl-L-methionine: Mg-protoporphyrin IX methyltransferase. J Bact 176: 5290–5296 (1994).

    PubMed  Google Scholar 

  9. Bollivar DW, Suzuki JY, Beatty JT, Dobrowolski J, Bauer CE: Directed mutational analysis of bacteriochlorophyll a biosynthesis in Rhodobacter capsulatus. J Mol Biol 237: 622–640 (1994).

    Article  PubMed  Google Scholar 

  10. Burke D, Alberti M, Hearst JE: bchFNBH bacteriochlorophyll synthesis genes of Rhodobacter capsulatus and identification of the third subunit of light-independent protochlorophyllide reductase in bacteria and plants. J Bact 175: 2414–2422 (1993).

    PubMed  Google Scholar 

  11. Choquet Y, Rahire M, Girard-Bascoe J, Erickson J, Rochaix J-D: A chloroplast gene is required for the light-independent accumulation of chlorophyll in Chlamydomonas reinhardtii. EMBO 11: 1697–1704 (1992).

    PubMed  Google Scholar 

  12. Fujita Y, Takahashi Y, Shonai F, Ogura Y, Matsubara H: Cloning, nucleotide sequences and differential expression of the nifH and nifH-like (frxC) genes from the filamentous nitrogen-fixing cyanobacterium Plectonema boryanum. Plant Cell Physiol 32: 1093–1106 (1991).

    Google Scholar 

  13. Fujita Y, Takahashi Y, Chuganji M, Matsubara H: The nifH-like (frxC) gene is involved in the biosynthesis of chlorophyll in the filamentous cyanobacterium Plectonema boryanum. Plant Cell Physiol 33: 81–92 (1992).

    Google Scholar 

  14. Fujita Y, Matsumoto H, Takahashi Y, Matsubara H: Identification of a nifDK-like gene (ORF467) involved in the biosynthesis of chlorophyll in the cyanobacterium Plectonema boryanum Plant Cell Physiol 34: 305–314 (1993).

    PubMed  Google Scholar 

  15. Fujita Y, Hase T: The third gene chlB, essential for the light-independent reduction of protochlorophyllide in the cyanobacterium Plectonema boryanum. Annual Meeting and 35th Symposia of the Japanese Society of Plant Physiology, March 28–30, 1995, Matsue, Japan, Abstract 361 (2pE0).

  16. Gibson LCD, Hunter CN: The bacteriochlorophyll biosynthesis gene, bchM of Rhodobacter sphaeroides encodes S-adenosyl-l-methionine: Mg-protoporphyrin IX transferase. FEBS Lett 352: 127–130 (1994).

    Article  PubMed  Google Scholar 

  17. Gibson LCD, Willows RD, Kannangara CG, Wettstein DV, Hunter CN: Magnesium-protoporphyrin chelatase of Rhodobacter sphaeroides: reconstitution of activity by combining the products of the bchH, -I and -D genes expressed in Escherichia coli Proc Natl Acad Sci USA 92: 1941–1944 (1995).

    PubMed  Google Scholar 

  18. Gribskov M, Devereaux J, Burgess J: The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression. Nucl Acids Res 12: 539–547 (1984).

    PubMed  Google Scholar 

  19. Hinchigeri SB, Nelson DW, Richards WR: Purification and reaction mechanism of S-adenosyl-lmethionine: magnesium protoporphyrin IX methyltransferase from Rhodopseudomonas sphaeroides. Photosynthetica 18: 168–178 (1984).

    Google Scholar 

  20. Hinchigeri SB, Richards WR: The reaction mechanism of S-adenosyl-l-methionine:magnesium protoporphyrin IX methyltransferase from Euglena gracilis. Photosynthetica 16: 554–560 (1982).

    Google Scholar 

  21. Hinchigeri SB, Chan JCS, Richards WR: Purification of S-adenosyl-l-methionine:magnesium protoporphyrin IX methyltransferase by affinity chromatography. Photosynthetica 15: 351–359 (1981).

    Google Scholar 

  22. Hudson A, Carpenter R, Doyle S, Coen ES: Olive: a key gene required for chlorophyll biosynthesis in Antirrhinum majus. EMBO J 12: 3711–3719 (1993).

    PubMed  Google Scholar 

  23. Jones OTG: Biosynthesis of porphyrins, hemes and chlorophylls. In: Clayton RK, Sistrom WR (eds) The Photosynthetic Bacteria, pp. 751–777. Plenum Press, New york (1978).

    Google Scholar 

  24. Koncz C, Mayerhofer R, Koncz-Kalman Z, Nawrath C, Reiss B, Rédei GP, Schell J. Isolation of a gene encoding a novel chloroplast protein by T-DNA tagging in Arabidopsis thaliana. EMBO J 9: 1337–1346 (1990).

    PubMed  Google Scholar 

  25. Li J, Goldschmidt-Clermont M, Timko M: Chloroplastencoded chlB is required for light-independent protochlorophyllide reductase activity of Chlamydomonas reinhardtii. Plant Cell 5: 1817–1829 (1993).

    Article  PubMed  Google Scholar 

  26. Lidholm J, Gustafsson P: Homologues of the green algal gidA gene and the liverwort frxC gene are present on the chloroplast genomes of conifers. Plant Mol Biol 17: 787–798 (1991).

    PubMed  Google Scholar 

  27. Liu X-Q, Xu H, Huang C: Chloroplast chlB gene is required for light-independent chlorophyll accumulation in Chlamydomonas reinhardtii. Plant Mol Biol 23: 297–308 (1993).

    PubMed  Google Scholar 

  28. Marrs BL: Mobilization of the genes for photosynthesis from Rhodopseudomonas capsulata by a promiscuous plasmid. J Bact 146: 1003–1012 (1981).

    PubMed  Google Scholar 

  29. Mattaj IW: RNA recognition: a family matter? Cell 73: 837–840 (1993).

    Article  PubMed  Google Scholar 

  30. Messing J, Vieira J: A new pair of M13 vectors for selecting either strand of double-digest restriction fragments. Gene 31: 269–273 (1982).

    Article  Google Scholar 

  31. Mulligan ME, Jackman DM, Murphy ST: Heterocystforming filamentous cyanobacteria encode proteins that resemble eukaryotic RNA-binding proteins of the RNP family. J Mol Biol 235: 1162–1170 (1994).

    Article  PubMed  Google Scholar 

  32. Ogura Y, Takemura M, Oda K, Yamata K, Ohta E, Fukuzawa H, Ohyama K. Cloning and nucleotide sequence of a frxC-ORF469 gene cluster of Synechocystis PCC6803: conservation with liverwort chloroplast frxC-ORF465 and nif operon. Biosci Biotechnol Biochem 56: 788–793 (1992).

    PubMed  Google Scholar 

  33. Orsat B, Monfort A, Chatellard P, Stutz E: Mapping and sequencing of an actively transcribed Euglena gracilis chloroplast gene (ccsA) homologous to the Arabidopsis thaliana nuclear gene cs (ch-42). FEBS Lett 303: 181–184 (1992).

    Article  PubMed  Google Scholar 

  34. Prentki P, Krisch HM: In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29: 303–313 (1984).

    Article  PubMed  Google Scholar 

  35. Suzuki JY, Bauer CE: Light-independent chlorophyll biosynthesis: Involvement of the chloroplast gene chlL(frxC). Plant Cell 4: 929–940 (1992).

    Article  PubMed  Google Scholar 

  36. Suzuki JY, Bauer CE: A prokaryotic origin for light-dependent chlorophyll biosynthesis of plants. Proc Natl Acad Sci USA 92: 3749–3753 (1995).

    PubMed  Google Scholar 

  37. Suzuki JY, Bauer CE: Altered monovinyl and divinyl protochlorophyllide pools in the bchJ mutant of Rhodobacter capsulatus: possible monovinyl substrate discrimination of light-independent protochlorophyllide reductase. J Biol Chem 270: 3732–3740 (1995).

    Article  PubMed  Google Scholar 

  38. Taylor DP, Cohen SN, Clark WG, Marrs BL: Alignment of the genetic and restriction maps of the photosynthetic region of the Rhodopseudomonas capsulata chromosome by a conjugation-mediated marker rescue technique. J Bact 154: 580–590 (1983).

    PubMed  Google Scholar 

  39. Tsudzuki J, Nakashima K, Tsudzuki T, Hiratsuka J, Shibata M, Wakasugi T, Sugiura M: Chloroplast DNA of black pine retains a residual inverted repeat lacking rRNA genes: nucleotide sequences of trnQ, trnK, psbA, trnI and trnH and the absence of rsp16. Mol Gen Genet 232: 206–214 (1992).

    PubMed  Google Scholar 

  40. Walker CJ, Mansfield KE, Smith KM, Castelfranco PA: Incorporation of atmospheric oxygen into the carbonyl functionality of the protochlorophyllide isocyclic ring. Biochem J 257: 599–602 (1989).

    PubMed  Google Scholar 

  41. Wu G, Williams HD, Zamanian M, Gibson F, Poole RK: Isolation and characterization of Escherichia coli mutants affected in aerobic respiration: the cloning and nucleotide sequence of ubiG. Identification of an S-adenosylmethionine-binding motif in protein, RNA, and small molecule methyltransferases. J Gen Microbiol 138: 2101–2112 (1992).

    PubMed  Google Scholar 

  42. Yang ZY, Bauer CE: Rhodobacter capsulatus genes involved in early steps of the bacteriochlorophyll biosynthetic pathway. J. Bact [1]: 619–629 (1990).

    Google Scholar 

  43. Young DA, Bauer CE, Williams JC, Marrs BL: Genetic evidence for superoperonal organization of genes for photosynthetic pigments and pigment-binding proteins in Rhodobacter capsulatus. Mol Gen Genet 218: 1–12 (1989).

    Article  PubMed  Google Scholar 

  44. Yildiz FH, Gest H, Bauer CE: Conservation of the photosynthesis gene cluster in Rhodospirillum centenum Mol Microbiol 6: 2683–2691 (1992).

    PubMed  Google Scholar 

  45. Zsebo KM, Hearst JE: Genetic physical mapping of a photosynthetic gene cluster from R. capsulata. Cell 37: 937–947 (1984).

    Article  PubMed  Google Scholar 

  46. Zucconi AP and Beatty JT (1988) Posttranscriptional regulation by light of the steady-state levels of mature B880–850 light harvesting complexes in Rhodobacter capsulatus. J Bacteriol 170: 877–882 (19[!]).

    PubMed  Google Scholar 

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Author notes

  1. Craig A. Smith

    Present address: Loyola University Medical School, 60611, Chicago, IL, USA

  2. Jon Y. Suzuki

    Present address: Center for Gene Research, Nagoya University, Furo-cho, Chikusa-ku, 464-01, Nagoya, Japan

Authors and Affiliations

  1. Department of Biology, Indiana University, Jordan Hall, 47405, Bloomington, IN, USA

    Craig A. Smith, Jon Y. Suzuki & Carl E. Bauer

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Smith, C.A., Suzuki, J.Y. & Bauer, C.E. Cloning and characterization of the chlorophyll biosynthesis gene chlM from Synechocystis PCC 6803 by complementation of a bacteriochlorophyll biosynthesis mutant of Rhodobacter capsulatus . Plant Mol Biol 30, 1307–1314 (1996). https://doi.org/10.1007/BF00019561

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  • DOI: https://doi.org/10.1007/BF00019561

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