Abstract
The N-terminal amino acid sequence of a 42.5 kDa subunit of the NADH: ubiquinone oxidoreductase (complex I) from potato has been determined by direct protein sequencing. The sequence was found to be homologous to that of the nuclear-encoded 49 kDa complex I subunit of bovine and Neurospora mitochondria and to the sequence deduced from the mitochondrial nad7 gene identified in the mitochondrial (mt) DNA of tryp anosomes and the moss Marchantia. An oligonucleotide probe derived from the potato N-terminal protein sequence hybridized only to the plant mtDNA. Immunoprecipitation of in-organello 35S-labelled potato and wheat mitochondrial translation products with an antibody directed against the Neurospora 49 kDa complex I subunit indicates that at least in these plants the NAD7 protein is synthesized within the organelle. Comparisons of genomic, cDNA and protein sequences of the 5′ coding region reveal three codons that are changed by RNA-editing and confirm translation of the edited transcripts in plant mitochondria. The NAD7 protein appears to undergo post-translational processing since the N-terminal methionine residue is absent from the mature mitochondrial protein.
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References
Attardi G, Schatz G (1988) Biogenesis of mitochondria. Annu Rev Cell Biol 4:289–333
Bonen L, Williams K, Bird S, Wood C (1994) The NADH dehydrogenase subunit 7 gene is interrupted by four, group II introns in the wheat mitochondrial genome. Mol Gen Genet 244:81–89
Braun HP, Schmitz K (1992) Affinity purification of cytochrome c reductase from potato mitochondria. Eur J Biochem 208:761–767
Braun HP, Schmitz K (1993) Purification and sequencing of cytochrome b from potato reveals methionine cleavage of a mitochondrially encoded protein. FEBS Lett 316:128–132
Begu D, Graves PV, Domec C, Arselin G, Litvak S, Araya A (1990) RNA editing of wheat mitochondrial ATP synthase subunit 9: direct protein and cDNA sequencing. Plant Cell 2:1283–1290
Chomczynski P, Sacchi A (1987) Single-step method of RNA isolation by guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–161
Covello PS, Gray MW (1989) RNA editing in plant mitochondria. Nature 341:592–597
Dell'Orto P, Moenne A, Graves PV, Jordana X (1993) The potato mitochondrial ATP synthase subunit 9: gene structure, RNA editing and partial protein sequence. Plant Sci 88:45–53
Fearnley IM, Runswick MJ, Walker JE (1989) A homologue of the nuclear encoded 49 kD subunit of the bovine mitochondrial NADH-ubiquinone reductase is coded in chloroplast DNA. EMBO J 8:665–672
Graves PV, Begu D, Velours J, Neau E, Belloc F, Litvak S, Araya A (1990) Direct protein sequencing of wheat mitochondrial ATP synthase subunit 9 confirms RNA editing in plants. J Mol Biol 214:1–6
Gualberto JM, Lamattina L, Bonnard G, Weil J-H, Grienenberger JM (1989) RNA editing in wheat mitochondria results in conservation of protein sequences. Nature 341:660–662
Gualberto JM, Bonnard G, Lamattina L, Grienenberger JM (1991) Expression of the wheat mitochondrial nad3-rps12 transcription unit: correlation between editing and mRNA maturation. Plant Cell 3:1109–1120
Herz U, Schröder W, Lidell A, Leaver CJ, Brennicke A, Grohmann L (1994) Purification of the NADH: Ubiquinone oxidoreductase (complex I) of the respiratory chain from the inner mitochondrial membrane of Solanum tuberosum. J Biol Chem 269:2263–2269
Hard F-U, Neupert W (1990) Protein sorting to mitochondria: evolutionary conservations of folding and assembly. Science 247:930–938
Hiesel R, Wissinger B, Schuster W Brennicke A (1989) RNA editing in plant mitochondria. Science 246:1632–1634
Hofhaus G, Weiss H, Leonhard K (1991) Electron microscopic analysis of the peripheral and membrane parts of mitochondrial NADH dehydrogenase (complex I). J Mol Biol 221:1027–1043
Koslowsky DJ, Bhat GJ, Perrollaz AL, Feagin JE, Stuart K (1990) The MURF3 gene of T. brucei contains multiple domains of extensive editing and is homologous to a subunit of NADH dehydrogenase. Cell 62:901–911
Leaver CJ, Hack E, Forde BG (1983) Protein synthesis by isolated plant mitochondria. Methods Enzymol 97:475–486
Leterme S, Boutry M (1993) Purification and preliminary characterization of mitochondrial complex I (NADH-ubiquinone reductase) from broad bean (Vicia faba). Plant Physiol 102:435–443
Macfarlane JL, Wahleithner JA, Wolstenholme DR (1990) A broad bean mitochondrial atp6 gene with an unusually simple, non-conserved 5′ region. Curr Genet 18:87–91
Maeshima M, Nakagawa T Asahi T (1989) N-terminal amino acid sequence and processing site of sweet potato cytochrome c oxidase subunit II. Plant Cell Physiol 30:1187–1188
Oda K, Yamato K, Ohta E, Nakamura Y, Takemura M, Nozato N, Akashi K, Kanege T, Ogura Y, Kohchi T, Ohyama K (1992) Gene organization deduced from the complete sequence of the liverwort Marchantia polymorpha mitochondrial DNA. J Mol Biol 223:1–7
Preis D, van der Pas JC, Nehls U, Röhlen DA, Sackmann U, Jahnke U, Weiss H (1990) The 49 kDa subunit of NADH: ubiquinone reductase (complex I) from Neurospora crassa mitochondria: primary structure of the gene anf the protein. Curr Genet 18:59–64
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Schuster W, Hiesel R, Wissinger B, Schobel W, Brennicke A (1989) Isolation and analysis of plant mitochondria and their genomes. In: Shaw CH (ed) Plant molecular biology. IRL Press, Oxford, Washington DC, pp 79–102
Schuster W, Wissinger B, Unseld M, Brennicke A (1990) Transcripts of the NADH dehydrogenase subunit 3 gene are differentially edited in Oenothera mitochondria. EMBO J 9:263–269
Shinozaki K, Ohme M, Tanaka M, Wakasugi T, Hayashida N, Matsubayashi T, Zaita N, Chunwongse 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 (1986) The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression. EMBO J 5:2043–2049
Souza AE, Myler PJ, Stuart K (1992) Maxicircle CR1 transcripts of Trypanosoma brucei are edited and developmentally regulated and encode a putative iron-sulphur protein homologous to an NADH dehydrogenase subunit. Mol Cell Biol 12:2100–2107
Walker JE (1992) The NADH: ubiquinone oxidoreductase (complex I) of respiratory chains. Q Rev Biophys 25:253–324
Weiss H, Friedrich T, Hofhaus G, Preis D (1991) The respiratory chain NADH dehydrogenase (complex I) of mitochondria. Eur J Biochem 197:563–576
Wissinger B, Schuster W, Brennicke A (1992) Regenerating good sense: RNA editing and trans-splicing in plant mitochondria. Trends Genet 8:322–328
Yang AJ, Mulligan RM (1991) RNA editing intermediates of cox2 transcripts in maize mitochondria. Mol Cell Biol 11:4278–4281
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Gäbler, L., Herz, U., Brennicke, A. et al. The 42.5 kDa subunit of the NADH: ubiquinone oxidoreductase (complex I) in higher plants is encoded by the mitochondrial nad7 gene. Molec. Gen. Genet. 244, 33–40 (1994). https://doi.org/10.1007/BF00280184
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DOI: https://doi.org/10.1007/BF00280184