Abstract
cDNA and genomic clones encoding the β subunit of mitochondrial ATP synthase from Chlamydomonas reinhardtii have been isolated using heterologous DNA probes from the photosynthetic bacterium Rhodospirillum rubrum. The protein encoded by the cDNA is 79–83% identical to corresponding proteins from higher-plant and mammalian mitochondria, and 75% identical to the R. rubrum protein. It contains both an N-terminal presequence and a unique C-terminal extension. The presequence, which is the first mitochondrial presequence determined in C. reinhardtii, is similar in structure to mitochondrial presequences from other organisms. As chloroplast presequences from C. reinhardtii also share features with mitochondrial presequences from other organisms (L.-G. Franzén et al., FEBS Lett 260 (1990) 165–168), this raises interesting questions about protein targeting to chloroplasts and mitochondria in C. reinhardtii. The possibility that the C-terminal extension is involved in targeting the protein to the mitochondrion is discussed. Southern blot analysis indicates that the protein is encoded by a single-copy gene.
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References
Amann R, Ludwig W, Schleifer KH: β-subunit of ATP-synthase: a useful marker for studying the phylogenetic relationship of eubacteria. J Gen Microbiol 134: 2815–2821 (1988).
Atteia A, de Vitry C, Pierre Y, Popot JL: Identification of mitochondrial proteins in membrane preparations of Chlamydomonas reinhardtii. J Biol Chem 267: 226–234 (1991).
Bankier AT, Barrell BG: Shotgun DNA sequencing. In: Flavell RA (ed) Techniques in Nucleic Acid Biochemistry B508, pp. 1–34. Elsevier, County Clare, Ireland/New York (1983).
Boutry M, Chua N-H: A nuclear gene encoding the beta subunit of the mitochondrial ATP synthase in Nicotiana plumbaginifolia. EMBO J 4: 2159–2165 (1985).
Boutry M, Nagy F, Poulsen C, Aoyagi K, Chua N-H: Targeting of bacterial chloramphenicol acetyltransferase to mitochondria in transgenic plants. Nature 328: 340–342 (1987).
Boutry M, Chaumont F, Leterme S, Castro Silva Filho M: Deciphering mitochondrial targeting presequences. In: Abstracts of the Fourth International Workshop on Plant Mitochondria, Ithaca, NY (1990).
Cozens AL, Walker JE: The organization and sequence of the genes for ATP synthase subunits in the cyanobacterium Synechococcus 6301. Support for an endosymbiotic origin of chloroplasts. J Mol Biol 194: 359–383 (1987).
Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 12: 387–395 (1984).
Docherty K, Steiner DF: Posttranslational proteolysis in polypeptide hormone biosynthesis. Annu Rev Physiol 44: 625–638 (1982).
Falk G, Hampe A, Walker JE: Nucleotide sequence of the Rhodospirillum rubrum atp operon. Biochem J 228: 391–407 (1985).
Farrel PJ, Deininger PL, Bankier A, Barrel L: Homologous upstream sequences near Epstein-Barr virus promoters. Proc Natl Acad Sci USA 80: 1565–1569 (1983).
Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).
Franzén L-G, Frank G, Zuber H, Rochaix J-D: Isolation and characterization of cDNA clones encoding Photosystem I subunits with molecular masses 11.0, 10.0 and 8.4 kDa from Chlamydomonas reinhardtii. Mol Gen Genet 219: 137–144 (1989).
Franzén L-G, Frank G, Zuber H, Rochaix J-D: Isolation and characterization of cDNA clones encoding the 17.9 and 8.1 kDa subunits of Photosystem I from Chlamydomonas reinhardtii. Plant Mol Biol 12: 463–474 (1989).
Franzén L-G, Rochaix J-D, von Heijne G: Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii share features with both mitochondrial and higher plant chloroplast presequences. FEBS Lett 260: 165–168 (1990).
Fukuzawa H, Fujiwara S, Yamamoto Y, Dionisio-Sese ML, Miyachi S: cDNA cloning, sequence and expression of carbonic anhydrase in Chlamydomonas reinhardtii: Regulation by environmental CO2 concentration. Proc Natl Acad Sci USA 87: 4383–4387 (1990).
Futai M, Noumi T, Maeda M: ATP synthase (H+ ATPase): Results by combined biochemical and molecular biological approaches. Annu Rev Biochem 58: 111–136 (1989).
Goldschmidt-Clermont M, Rahire M: Sequence, evolution and differential expression of the two genes encoding variant small subunits of ribulose bisphospate carboxylase/oxygenase in Chlamydomonas reinhardtii. J Mol Biol 191: 421–432 (1986).
Gray MW: The evolutionary origins of organelles. Trends Genet 5: 294–299 (1989).
Harris DA: The interaction of coupling factor ATPases with nucleotides. Biochim Biophys Acta 463: 245–273 (1978).
Hawthorne CA, Brusilow WSA: Sequence of the genes for the β and ε subunits of the ATP synthase of Bacillus megaterium QM B1551. Biochem Biophys Res Commun 151: 926–931 (1988).
Hurt EC, Soltanifar N, Goldschmidt-Clermont M, Rochaix JD, Schatz G: The cleavable pre-sequence of an imported chloroplast protein directs attached polypeptides into yeast mitochondria. EMBO J 5: 1343–1350 (1986).
Imbault P, Wittemer C, Johanningmeier U, Jacobs JD and Howell SH: Structure of the Chlamydomonas reinhardtii cabII-1 gene encoding a chlorophyll a/b-binding protein. Gene 73: 397–407 (1988).
Kozak M: An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucl Acids Res 15: 8125–8148 (1987).
Mayfield SP, Rahire M, Frank G, Zuber H, Rochaix J-D: Expression of the nuclear gene encoding oxygenevolving enhancer protein 2 is required for high levels of photosynthetic oxygen evolution in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 84: 749–753 (1987).
Mayfield SP, Schirmer-Rahire M, Frank G, Zuber H, Rochaix J-D: Analysis of the genes of the OEE1 and OEE3 proteins of the photosystem II complex from Chlamydomonas reinhardtii. Plant Mol Biol 12: 683–693 (1989).
Merchant S, Hill K, Kim JH, Thompson J, Zaitlin D, Bogorad L: Isolation and characterization of a complementary DNA clone for an algal pre-apoplastocyanin. J Biol Chem 265: 12372–12379 (1990).
Mitchell P: Coupling of phosphorylation to electron and hydrogen transfer by a chemi-osmotic type of mechanism. Nature 191: 144–148 (1961).
Mitchell P: A chemiosmotic molecular mechanism for proton-translocating adenosine triphosphatases. FEBS Lett 43: 189–194 (1974).
Needleman SB, Wunsch CD: A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol 48: 443–453 (1970).
Ohta S, Kagawa Y: Human F1-ATPase: Molecular cloning of cDNA for the beta subunit. J Biochem 99: 135–141 (1986).
Rochaix J-D: Molecular genetics of chloroplasts and mitochondria in the unicellular green alga Chlamydomonas. FEMS Microbiol Rev 46: 13–34 (1987).
Rochaix J-D, Mayfield S, Goldschmidt-Clermont M, Erickson J: Molecular Biology of Chlamydomonas. In: Shaw CH (ed) Plant Molecular Biology, pp. 253–275. IRL Press, Oxford (1988).
Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Saraste M, Gay NJ, Eberle A, Runswick MJ, Walker JE: The atp operon: Nucleic acid sequence of the genes for the gamma, beta and epsilon subunits of Escherichia coli ATP synthase. Nucl Acids Res 9: 5287–5296 (1981).
Shinozaki K, Deno H, Kato A, Sugiura M: Overlap and cotranscription of the genes for the beta and epsilon subunits of tobacco chloroplast ATPase. Gene 24: 147–155 (1983).
Silflow CD, Chisholm RL, Conner TW, Ranum LPW: The two alpha tubulin genes of Chlamydomonas reinhardtii code for slightly different proteins. Mol Cell Biol 5: 2389–2398 (1985).
Staden R: Automation of the computer handling of gel reading data produced by the shotgun method of DNA sequencing. Nucl Acids Res 10: 4731–4751 (1982).
Staden R: An interactive graphics program for comparing and aligning nucleic acid and amino sequences. Nucl Acids Res 10: 2951–2961 (1982).
Staden R: Computer methods to locate genes and signals in nucleic acid sequences. In: Setlow YK, Hollaender A (ed) Genetic Engineering: Principles and Methods, vol 7, pp. 67–114. Plenum, New York/London (1985).
Takeda M, Vassarotti A, Douglas MG: Nuclear genes coding the yeast mitochondrial adenosine triphosphatase complex. J Biol Chem 260: 15458–15465 (1985).
Walker JE, Fearnley IM, Gay NJ, Gibson BW, Northrop FD, Powell SJ, Runswick MJ, Saraste M, Tybulewicz VLJ: Primary structure and subunit stoichiometry of F1-ATPase from bovine mitochondria. J Mol Biol 184: 677–701 (1985).
von Heijne G, Steppuhn J, Herrmann RG: Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem 180: 535–545 (1989).
Whelan J, Knorpp C, Glaser E: Sorting of precursor proteins between isolated leaf mitochondria and chloroplasts. Plant Mol Biol 14: 977–982 (1990).
Woessner JP, Gillham NW, Boynton JE: The sequence of the chloroplast atpB gene and its flanking sequence in Chlamydomonas reinhardtii. Gene 44: 17–28 (1986).
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Franzén, LG., Falk, G. Nucleotide sequence of cDNA clones encoding the β subunit of mitochondrial ATP synthase from the green alga Chlamydomonas reinhardtii: The precursor protein encoded by the cDNA contains both an N-terminal presequence and a C-terminal extension. Plant Mol Biol 19, 771–780 (1992). https://doi.org/10.1007/BF00027073
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DOI: https://doi.org/10.1007/BF00027073