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Primary structure of the plasma membrane H+-ATPase from the halotolerant alga Dunaliella bioculata

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Abstract

P-type ATPase-specific oligodeoxyribonucleotides were used to obtain a fragment of the H+-ATPase of the salt tolerant alga Dunaliella bioculata by polymerase chain reaction (PCR). This fragment served as a probe in screening a cDNA-library from this organism. The complete primary structure of the ATPase protein (DBPMA1) was deduced from sequencing a 4.7 kb cDNA clone. The protein shows highest homology to H+-ATPases from higher plants and fungi (43% identity, 67% similarity) but has a higher calculated molecular mass (123 kDa). The latter can be assigned mainly to an additional hydrophilic domain between transmembrane segments VI and VII and to an extended carboxyterminus. These unusual structural features of DBPMA1 are interpreted in terms of providing regulatory sites of the enzyme. Southern blot analysis suggests the presence of only a single copy of the gene in the haploid D. bioculata genome. To investigate the role of the H+-ATPase in the adaption of D. bioculata to different external NaCl concentrations, we employed northern blot analyses. The results indicate that the pmal transcript level of cells growing in salinities between 0.1 and 3 M NaCl is not directly correlated with the external salt concentration.

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References

  1. Bental M, Degani H, Avron M: 23Na-NMR studies of the intracellular sodium ion concentration in the halotolerant alga Dunaliella salina. Plant Physiol 87: 813–817 (1988).

    Google Scholar 

  2. Chang A, Slayman CW: Maturation of the yeast plasma membrane <H+>ATPase involves phosphorylation during intracellular transport. J Cell Biol 115: 289–295 (1991).

    Google Scholar 

  3. Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159 (1987).

    Google Scholar 

  4. Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995 (1984).

    Google Scholar 

  5. Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence analysis programs for the VAX. Nucl Acids Res 121: 387–395 (1984).

    Google Scholar 

  6. Ewing NN, Wimmers LE, Meyer DJ, Chetelat RT, Bennett AB: Molecular cloning of tomato plasma membrane H+-ATPase. Plant Physiol 94: 1874–1881 (1990).

    Google Scholar 

  7. Fickett JW: Recognition of protein coding regions in DNA sequences. Nucl Acids Res 10: 5303–5318 (1982).

    Google Scholar 

  8. Gilmour DJ, Kaaden R, Gimmler H: Vanadate inhibition of ATPases of Dunaliella parva in vitro and in vivo. J Plant Physiol 18: 111–126 (1985).

    Google Scholar 

  9. Hager KM, Mandala SM, Davenport JW, Speicher DW, Benz EJ, Slayman CW: Amino acid sequence of the plasma membrane ATPase of Neurospora crassa: deduction from genomic and cDNA sequences. Proc Natl Acad Sci USA 83: 7693–7697 (1986).

    Google Scholar 

  10. Harper JF, Surowy TK, Sussman MR: Molecular cloning and sequence of cDNA encoding the plasma membrane proton pump (H+-ATPase) of Arabidopsis thaliana. Proc Natl Acad Sci USA 86: 1234–8 (1989).

    Google Scholar 

  11. Harper JF, Sussman MR, Schaller GE, Putnam-Evans C, Charbonneau H, Harmon AC: A calcium-dependent protein kinase with a regulatory domain similar to calmodulin. Science 252: 951–954 (1991).

    Google Scholar 

  12. Hopp TP, Wood KR: Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci USA 78: 3824–3828 (1981).

    Google Scholar 

  13. Johansson F, Sommarin M, Larsson C: Fusicoccin activates the plasma membrane H+-ATPase by a mechanism involving the C-terminal inhibitory domain. Plant Cell 5: 321–327 (1993).

    Google Scholar 

  14. Katz A, Bental M, Degani H, Avron M: In vivo pH regulation by a Na+/H+ antiporter in the halotolerant alga Dunaliella salina. Plant Physiol 96: 110–115 (1991).

    Google Scholar 

  15. Katz A, Kaback HR, Avron M: Na+/H+ antiport in isolated plasma membrane vesicles from the halotolerant alga Dunaliella salina. FEBS Lett 202: 141–144 (1986).

    Google Scholar 

  16. Katz A, Pick U, Avron M: Characterization and reconstitution of the Na+/H+ antiporter from the plasma membrane of the halotolerant alga Dunaliella. Biochim Biophys Acta 983: 9–14 (1989).

    Google Scholar 

  17. Kolarov J, Kulpa J, Baijot M, Goffeau A: Characterization of a protein serine kinase from yeast plasma membrane. J Biol Chem 263: 10613–10619 (1988).

    Google Scholar 

  18. Kozak M: An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucl Acids Res 15: 8135–8148 (1987).

    Google Scholar 

  19. LaRoche J, Bennett J, Falkowski PG: Characterization of a cDNA encoding for the 28.5-kDa LHCII apoprotein from the unicellular marine chlorophyte, Dunaliella tertiolecta. Gene 95: 165–171 (1990).

    Google Scholar 

  20. Lütcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA: Selection of AUG initiation codons differs in plants and animals. EMBO J 6: 43–48 (1987).

    Google Scholar 

  21. Nakamoto RK, Rao R, Slayman CW: Transmembrane segments of the P-type cation-transporting ATPases. A comparative study. Ann N Y Acad Sci 574: 165–179 (1989).

    Google Scholar 

  22. Niu XM, Zhu JK, Narasimhan ML, Bressan RA, Hasegawa PM: Plasma membrane H+-ATPase gene expression is regulated by NaCl in cells of the halophyte Atriplex nummularia L. Planta 190: 433–438 (1993).

    Google Scholar 

  23. Oren-Shamir M, Pick U, Avron M: Plasma membrane potential of the alga Dunaliella and its relation to osmoregulation. Plant Physiol 93: 403–408 (1990).

    Google Scholar 

  24. Palmgren MG, Sommarin M, Serrano R, Larsson C: Identification of an autoinhibitory domain in the C-terminal region of the plant plasma membrane H(+)-ATPase. J Biol Chem 266: 20470–20475 (1991).

    Google Scholar 

  25. Pick U: ATPases and ion transport in Dunaliella. In: Avron M, Benamotz A (eds) Dunaliella: Physiology, Biochemistry, and Biotechnology, pp. 63–97. CRC Press, Boca Raton, FL (1992).

    Google Scholar 

  26. Pick U, Karni L, Avron M: Determination of ion content and ion fluxes in the halotolerant alga Dunaliella salina. Plant Physiol 81: 92–96 (1986).

    Google Scholar 

  27. Popova LG, Balnokin YV: H+-translocating ATPase and Na+/H+ antiport activities in the plasma membrane of the marine alga Platymonas viridis. FEBS Lett 309: 333–336 (1992).

    Google Scholar 

  28. Portillo F, deLarrinoa IF, Serrano R: Deletion analysis of yeast plasma membrane H+-ATPase and identification of a regulatory domain at the carboxyl-terminus. FEBS Lett 247: 381–385 (1989).

    Google Scholar 

  29. Roberts DM, Harmon AC: Calcium-modulated proteins: targets of intracellular calcium signals in higher plants. Ann Rev Plant Physiol Plant Mol Biol 43: 375–414 (1992).

    Google Scholar 

  30. Schaller GE, Sussman MR: Phosphorylation of the plasma-membrane H-ATPase of oat roots by a calcium-stimulated protein kinase. Planta 173: 509–518 (1988).

    Google Scholar 

  31. Sekler I, Gläser HU, Pick U: Characterization of a plasma membrane H+-ATPase from the extremely acidophilic alga Dunaliella acidophila. J Membrane Biol 121: 51–57 (1991).

    Google Scholar 

  32. Sekler I, Pick U: Purification and properties of a plasma membrane H+-ATPase from the extremely acidophilic alga Dunaliella acidophila. Plant Physiol 101: 1055–1061 (1993).

    Google Scholar 

  33. Sekler I, Weiss M, Pick U: Activation of the Dunaliella acidophila plasma membrane H+-ATPase by trypsin cleavage of a fragment that contains a phosphorylation site. Plant Physiol 105: 1125–1132 (1994).

    Google Scholar 

  34. Serrano R: Structure and function of plasma membrane ATPase. Annu Rev Plant Physiol Plant Mol Biol 40: 61–94 (1989).

    Google Scholar 

  35. Serrano R, Kielland Brandt MC, Fink GR: Yeast plasma membrane ATPase is essential for growth and has homology with (Na++K+), K+- and Ca2+-ATPases. Nature 319: 689–693 (1986).

    Google Scholar 

  36. Serrano R, Portillo F, Monk BC, Palmgren MG: The regulatory domain of fungal and plant plasma membrane H+-ATPase. Acta Physiol Scand 146: 131–136 (1992).

    Google Scholar 

  37. Siflow CD, Chisholm RL, Conner TW, Raneem LPW: The two alpha-tubulin genes of Chlamydomonas reinhardtii code for slightly different proteins. Mol Cell Biol 5: 2389–2398 (1985).

    Google Scholar 

  38. Smahel M, Hamann A, Gradmann D: The prime plasmalemma ATPase of the halophilic alga Dunaliella bioculata: purification and characterization. Planta 181: 496–504 (1990).

    Google Scholar 

  39. Suzuki YS, Wang YL, Takemoto JY: Syringomycin-stimulated phosphorylation of the plasma membrane H+-ATPase from red beet storage tissue. Plant Physiol 99: 1314–1320 (1992).

    Google Scholar 

  40. Wada M, Takano M, Kasamo K: Nucleotide sequence of a complementary DNA encoding plasma membrane H+-ATPase from rice (Oryza sativa L). Plant Physiol 99: 794–795 (1992).

    Google Scholar 

  41. Wada M, Urayama O, Satoh S, Hara Y, Ikawa Y, Fujii T: A marine algal Na+-activated ATPase possesses an immunologically identical epitope to Na+, K+-ATPase. FEBS Lett 309: 272–274 (1992).

    Google Scholar 

  42. Weiss M, Sekler I, Pick U: Characterization of soluble and membrane-bound forms of a vanadate-sensitive ATPase from plasma membranes of the halotolerant alga Dunaliella salina. Biochim Biophys Acta 974: 254–260 (1989).

    Google Scholar 

  43. Wolf AH, Kirsch M, Wiessner W: Nucleotide sequence of a cDNA encoding ribosomal protein L27 from Chlamydobotrys stellata. Plant Physiol 101: 1123 (1993).

    Google Scholar 

  44. Yuasa T, Muto S: Ca2+-dependent protein kinase from the halotolerant green alga Dunaliella tertiolecta: partial purification and Ca2+-dependent association of the enzyme to the microsomes. Arch Biochem Biophys 296: 175–182 (1992).

    Google Scholar 

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Authors and Affiliations

  1. Pflanzenphysiologisches Institut der Universität, Untere Karspüle 2, 37073, Göttingen, Germany

    Andreas H. Wolf & Dietrich Gradmann

  2. Department of Genetics, Yale School of Medicine, 333 Cedar Street, 06510, New Haven, CT, USA

    Carolyn W. Slayman

  3. Department of Cellular and Molecular Physiology, Yale School of Medicine, 333 Cedar Street, 06510, New Haven, CT, USA

    Carolyn W. Slayman

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  1. Andreas H. Wolf
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  2. Carolyn W. Slayman
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  3. Dietrich Gradmann
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Wolf, A.H., Slayman, C.W. & Gradmann, D. Primary structure of the plasma membrane H+-ATPase from the halotolerant alga Dunaliella bioculata . Plant Mol Biol 28, 657–666 (1995). https://doi.org/10.1007/BF00021191

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  • DOI: https://doi.org/10.1007/BF00021191

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