Skip to main content
Log in

Cloning and transcription analysis of the ndh(A-I-G-E) gene cluster and the ndhD gene of the cyanobacterium Synechocystis sp. PCC6803

  • Research Articles
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The plastid DNA of higher plants contains eleven reading frames that are homologous to subunits of the mitochondrial NADH-ubiquinone oxidoreductase (complex I). The genes are expressed, but a plastid NAD(P)H dehydrogenase has not yet been isolated and the function of the enzyme in plastid metabolism is unknown. Cyanobacteria also contain a NADH dehydrogenase that is homologous to the mitochondrial complex I. The enzyme is sensitive to rotenone and is located on the cytoplasmic and the thylakoid membrane. We report here the sequence of five subunits (ndhA,-I, G,-E and -D) of the NADH dehydrogenase from the unicellular cyanobacterium Synechocystis sp. PCC6803. As in plastid DNA, the genes ndh(A-I-G-E) are clustered and probably constitute an operon. The ndhD gene is associated with a gene encoding an iron-sulphur protein of photosystem I (psaC) as in plastid DNA. In contrast to the situation in plastids, psaC and ndhD are not cotranscribed but transcribed from opposite strands. The deduced amino acid sequence of the cyanobacterial polypeptides is more similar to the corresponding plastid (40–68% identity) than to the corresponding mitochondrial subunits (17–39% identity). Thus, the cyanobacterial NADH-dehydrogenase provides a prokaryotic model system which is more suitable to genetic analysis than the enzyme of plastids.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Anderson S, Bruijn MHL, Coulson AR, Eperon IC, Sanger F, Young IG: Complete sequence of bovine mitochondrial DNA. J Mol Biol 156: 683–717 (1982).

    PubMed  Google Scholar 

  2. Anderson SL, McIntosh L: Partial conservation of the 5′ ndhE-psaC-ndhD 3′ gene arrangement of chloroplasts in the cyanobacterium Synechocystis sp. PCC6803: implications for NDH-D function in cyanobacteria and chloroplasts. Plant Mol Biol 16: 487–499 (1991).

    PubMed  Google Scholar 

  3. Bennoun P: Evidence for a respiratory chain in the chloroplast. Proc Natl Acad Sci USA 79: 4342–4356 (1982).

    PubMed  Google Scholar 

  4. Berger S, Ellersiek U, Steinmüller K: Cyanobacteria contain a mitochondrial complex I-homologous NADH-dehydrogenases. FEBS Lett 286: 129–132 (1991).

    PubMed  Google Scholar 

  5. Böhm R, Sauter M, Böck A: Nucleotide sequence and expression of an operon in Escherichia coli coding for formate hydrogenylase components. Mol Microbiol 4: 231–243 (1990).

    PubMed  Google Scholar 

  6. Burger G, Werner S: The mitochondrial URF1 gene in Neurospora crassa has an intron that contains a novel type of URF. J Mol Biol 186: 231–242 (1985).

    PubMed  Google Scholar 

  7. Chomyn A, Cleeter MWJ, Ragan CI, Riley M, Doolittle RF, Attardi G: URF6, last unidentified reading frame of human mtDNA, codes for an NADH dehydrogenase subunit. Science 234: 614–618 (1986).

    PubMed  Google Scholar 

  8. Clayton DA: Replication and transcription of vertebrate mitochondrial DNA. Annu Rev Cell Biol 7: 453–478 (1991).

    PubMed  Google Scholar 

  9. deVries H, Alzner-DeWeerd B, Breitenberger CA, Chang DD, deJonge JC, TajBhandary UL: The E35 stopper mutant of Neurospora crassa: precise localization of deletion endpoints in mitochondrial DNA and evidence that the deleted DNA codes for a subunit of NADH dehydrogenase. EMBO J 5: 779–785 (1986).

    PubMed  Google Scholar 

  10. Dupuis A, Skehel JM, Walker JE: A homologue of a nuclear-coded iron-sulphur protein subunit of bovine mitochondrial complex I is encoded in chloroplast genomes. Biochemistry 30: 2954–2960 (1991).

    PubMed  Google Scholar 

  11. Dzelzkalns VA, Bogorad L: Stable transformation of the cyanobacterium Synechocystis sp. PCC6803 induced by UV irradiation. J. Bact 165: 964–971 (1986).

    PubMed  Google Scholar 

  12. Earley FGP, Patel SD, Ragan CI, Attardi G: Photolabelling of a mitochondrially encoded subunit of NADH dehydrogenase with [3H]dihydrorotenone. FEBS Lett 219: 108–113 (1987).

    PubMed  Google Scholar 

  13. Fearnley IM, Runswick MJ, Walker JE: A homologue of the nuclear coded 49 kd subunit of bovine mitochondrial NADH-ubiquinone reductase is coded in chloroplast DNA. EMBO J 8: 665–672 (1989).

    PubMed  Google Scholar 

  14. Friedrich T, Hofhaus G, Ise W, Nehls U, Schmitz B, Weiss H: A small isoform of NADH:ubiquinone oxidoreductase (complex I) without mitochondrially encoded subunits is made in chloramphenicol-treated Neurospora crassa. Eur J Biochem 180: 173–180 (1989).

    PubMed  Google Scholar 

  15. Godde D, Trebst A: NADH as electron donor for the photosynthetic membrane of Chlamydomonas reinhardtii. Arch Microbiol 127: 245–252 (1980).

    Google Scholar 

  16. Godde D: Evidence for a membrane-bound NADH-plastoquinone-oxidoreductase in Chlamydomonas reinhardtii CW-15. Arch Microbiol 131: 197–202 (1982).

    Google Scholar 

  17. Golbeck JH, Bryant DA: Photosystem I. Curr Top Bioenerget 16: 83–177 (1991).

    Google Scholar 

  18. Hiratsuka J, Shimada H, Whittier R, Ishibashi T, Sakamoto M, Mori M, Kondo C, Honji Y, Sun CR, Meng BY, Li YQ, Kanno A, Nishizawa Y, Hirai A, Shinozaki K, Sugiura M: The complete sequence of the rice (Oryza sativa) chloroplast genome: Intermolecular recombination between distinct tRNA genes accounts for a major plastid DNA inversion during the evolution of the cereals. Mol Gen Genet 217: 185–194 (1989).

    PubMed  Google Scholar 

  19. Kohchi T, Shirai H, Fukuzawa H, Sano T, Komano T, Umesono K, Inokuchi H, Ozeki H, Ohyama K: Structure and organization of Marchantia polymorpha chloroplast genome. IV; Inverted repeat and small single copy regions. J Mol Biol 203: 353–372 (1988).

    Article  PubMed  Google Scholar 

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

    Google Scholar 

  21. Masui R, Wakabayashi S, Matsubara H, Hatefi Y: The amino acid sequence of the 9 kDa polypeptide and partial amino acid sequence of the 20 kDa polypeptide of mitochondrial NADH:ubiquinone oxidoreductase. J Biochem 110: 575–582 (1991).

    PubMed  Google Scholar 

  22. Matsubayashi T, Wakasugi T, Shinozaki K, Yamaguchi-Shinozak K, Zaita N, Hadaka T, Meng BY, Ohto C, Tanaka M, Kato A, Maruyama T, Sugiura M: Six chloroplast genes (ndhA-F) homologous to human mitochondrial genes encoding components of the respiratory chain NADH dehydrogenase are actively expressed: Determination of the splice sites in ndhA and ndhB pre-mRNAs. Mol Gen Genet 210: 385–393 (1987).

    Article  PubMed  Google Scholar 

  23. Mazel D, Houmard J, Tandeau de Marsac N: A multigene family in Calothrix sp. PCC7601 encodes phycocyanin, major component of the cyanobacterial light harvesting antenna. Mol Gen Genet 211: 296–304 (1988).

    Article  Google Scholar 

  24. Meijer EM, Schuitenmaker MG, Boogerd FC, Wever R, Stouthamer AH: Effects induced by rotenone during aerobic growth of Paracoccus denitrificans in continuous culture. Arch Microbiol 119: 119–127 (1978).

    PubMed  Google Scholar 

  25. Mulligan B, Schultes N, Chen L, Bogorad L: Nucleotide sequence of a multiple-copy gene for the B protein of photosystem II of a cyanobacterium. Proc Natl Acad Sci USA 81: 2693–2697 (1984).

    Google Scholar 

  26. Nelson MA, Macino G: Structure and expression of the overlapping ND4L and ND5 genes of Neurospora crassa mitochondria. Mol Gen Genet 206: 307–317 (1987).

    Article  PubMed  Google Scholar 

  27. Ogawa T: A gene homologous to the subunit-2 of NADH dehydrogenase is essential to inorganic carbon transport of Synechocystis sp. PCC6803. Proc Natl Acad Sci USA 88: 4275–4279 (1991).

    PubMed  Google Scholar 

  28. Ohyama K, Fukuzawa H, Kohchi T, Shirai H, Sano T, Sano S, Umesono K, Shiki Y, Takeuchi M, Chang Z, Aota SI, Inokuchi H, Ozeki H: Chloroplast gene organization deduced from complete sequence of liverwort Marchantia polymorpha chloroplast DNA. Nature 322: 572–574 (1986).

    Google Scholar 

  29. Ohyama K, Kohchi T, Sano T, Yamada Y: Newly identified groups of genes in chloroplast. Trends Biochem Sci 13: 19–22 (1988).

    Article  PubMed  Google Scholar 

  30. Pilkington SJ, Skehel JM, Walker JE: The 30-kilodalton subunit of bovine mitochondrial complex I is homologous to a protein coded in chloroplast DNA. Biochemistry 30: 1901–1908 (1991).

    PubMed  Google Scholar 

  31. Preis D, van derPas JC, Nehls U, Röhlen DA, Sackmann U, Jahnke U, Weiss H: The 49 K subunit of NADH:ubiquinone reductase (complex I) from Neurospora crassa mitochondria: primary structure of the gene and the protein. Curr Genet 18: 59–64 (1990).

    Article  PubMed  Google Scholar 

  32. Ragan CI: Structure of NADH-ubiquinone reductase (Complex I). Curr Top Bioenerget 15: 1–36 (1987).

    Google Scholar 

  33. Reed KC, Mann DA: Rapid transfer of DNA from agarose gels to nylon membranes. Nucl Acids Res 18: 173–180 (1985).

    Google Scholar 

  34. Schantz R, Bogorad L: Maize chloroplast genes ndhD, ndhE, and psaC. Sequences, transcripts and transcript pools. Plant Mol Biol 11: 239–247 (1988).

    Google Scholar 

  35. Schwitzguébel JP, Palmer JM: Properties of mitochondria as a function of the growth stages of Neurospora crassa. J Bact 149: 612–619 (1982).

    PubMed  Google Scholar 

  36. Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chungwongse J, Obokata J, Yamaguchi-Shinozaki K, Ohto C, Torazawa K, Meng BY, Sugita M, Deno H, Kamogashira T, Yamada K, Kusuda J, Takaiwa F, Kato A, Tohdoh N, Shimada H, Sugiura M: The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J 5: 2043–2049 (1986).

    Google Scholar 

  37. Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chungwongse J, Obokata J, Yamaguchi-Shinozaki K, Ohto C, Torazawa K, Meng BY, Sugita M, Deno H, Kamogashira T, Yamada K, Kusuda J, Takaiwa F, Kato A, Tohdoh N, Shimada H, Sugiura M: The complete nucleotide sequence of the tobacco chloroplast genome. Plant Mol Biol Rep 4: 111–147 (1986).

    Google Scholar 

  38. Steinmüller K, Ley AC, Steinmetz AA, Sayre RT, Bogorad L: Characterization of the ndhC-psbG-ORF157/159 operon of maize plastid DNA and of the cyanobacterium Synechocystis sp. PCC6803. Mol Gen Genet 216: 60–69 (1986).

    Article  Google Scholar 

  39. Steinmüller K, Ellersiek U, Bogorad L: Deletion of the psbG1 gene of the cyanobacterium Synechocystis sp. PCC6803 leads to the activation of the cryptic psbG2 gene. Mol Gen Genet 226: 107–112 (1991).

    Article  PubMed  Google Scholar 

  40. Steinmüller K: Nucleotide sequence and expression of the ndhH gene of the cyanobacterium Synehocystis sp. PCC6803. Plan Mol Biol 18: 135–137 (1992).

    Google Scholar 

  41. Steinmüller K: Identification of a second psaC gene in the cyanobacterium Synechocystis sp. PCC6803 (Submitted).

  42. Takahashi Y, Shonai F, Fujita Y, Kohchi T, Ohyama K, Matsubara H: Structure of a co-transcribed gene cluster, ndh1-frxB-ndh6-ndh4L, cloned from the filamentous cyanobacterium Plectonema boryanum. Plant Cell Physiol 32: 969–981 (1991).

    Google Scholar 

  43. Tuschen G, Sackmann U, Nehls U, Haiker H, Buse G, Weiss H: Assembly of NADH:ubiquinone reductase (Complex I) in Neurospora mitochondria. J Mol Biol 213: 845–857 (1990).

    PubMed  Google Scholar 

  44. Umesono K, Inokuchi H, Shiki Y, Takeuchi M, Chang Z, Fukuzawa H, Kohchi T, Shirai H, Ohyama K, Ozeki H: Structure and organization of Marchantia polymorpha chloroplast genome. II. Gene organization of the large single copy region from rps' 12 to atpB. J Mol Biol 203: 299–331 (1988).

    Article  PubMed  Google Scholar 

  45. Videira A, Tropschug M, Werner S: Primary structure and expression of a nuclear-coded subunit of complex I homologous to proteins specified by the chloroplast genome. Biochem Biophys Res Comm 171: 1168–1174 (1990).

    PubMed  Google Scholar 

  46. Weiss H, Friedrich T, Hofhaus G, Preis D: The respiratory-chain NADH dehydrogenase (complex I) of mitochondria. Eur J Biochem 197: 563–576 (1991).

    PubMed  Google Scholar 

  47. Wu M, Nie ZQ, Yang J: The 18 kD protein that binds to the chloroplast DNA replicative origin is an iron-sulfur protein related to a subunit of NADH dehydrogenase. Plant Cell 1: 551–557 (1989).

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ellersiek, U., Steinmüller, K. Cloning and transcription analysis of the ndh(A-I-G-E) gene cluster and the ndhD gene of the cyanobacterium Synechocystis sp. PCC6803. Plant Mol Biol 20, 1097–1110 (1992). https://doi.org/10.1007/BF00028896

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00028896

Key words

Navigation