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Protein phosphatases in higher plants: multiplicity of type 2A phosphatases in Arabidopsis thaliana

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Abstract

Two DNA fragments, AP-1 and AP-2, encoding amino acid sequences closely related to Ser/Thr protein phosphatases were amplified from Arabidopsis thaliana genomic DNA. Fragment AP-1 was used to screen. A. thaliana cDNA libraries and several positive clones were isolated. Clones EP8a and EP14a were sequenced and found to encode almost identical proteins (97% identity). Both proteins are 306 amino acids in length and are very similar (79–80% identity) to the mammalian isotypes of the catalytic subunit of protein phosphatase 2A. Therefore, they have been designated PP2A-1 and PP2A-2. A third cDNA clone, EP7, was isolated and sequenced. The polypeptide encoded (308 amino acids, lacking the initial Met codon) is 80% identical with human phosphatases 2A and was named PP2A-3. The PP2A-3 protein is extremely similar (95% identity) to the predicted protein from a cDNA clone previously found in Brassica napus. Southern blot analysis of genomic DNA using AP-1 and AP-2 probes, as well as probes derived from clones EP7, EP8a and EP14a strongly indicates that at least 6 genes closely related to type 2A phosphatases are present in the genome of A. thaliana. Northern blot analysis using the same set of probes demonstrates that, at the seedling stage, the mRNA levels for PP2A-1, PP2A-3 and the gene containing the AP-1 sequence are much higher than those of PP2A-2 and AP-2. These results demonstrate that a multiplicity of type 2A phosphatases might be differentially expressed in higher plants.

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References

  1. Ariño J, Woon CW, Brautigan DL, Miller TB, Johnson GL: Human liver phosphatase 2A: cDNA and amino acid sequence of two catalytic subunit isotypes. Proc Natl Acad Sci USA 85: 4252–4256 (1988).

    PubMed  Google Scholar 

  2. Budde RJA, Randall DD: Protein kinases in higher plants. In: Morre DJ, Boss WF, Loewus FA (eds) Inositol Metabolism in Plants, pp. 351–367. Wiley-Liss, New York (1990).

    Google Scholar 

  3. Budde RJA, Chollet R: Regulation of enzyme activity in plants by reversible phosphorylation. Physiol Plant 72: 435–439 (1988).

    Google Scholar 

  4. Carter PJ, Nimmo HG, Fewson CA, Wilkins MB: Bryophyllum fedtschenkol protein phosphatase type 2A can desphosphorylate phosphoenolpyruvate carboxylase. FEBS Lett 263: 233–236 (1990).

    Article  Google Scholar 

  5. Cohen P: The structure and regulation of protein phosphatases. Annu Rev Biochem 58: 453–508 (1989).

    Article  PubMed  Google Scholar 

  6. Cohen P: pp. 230–235. In: Nishizuka Y et al. (eds) The Biology and Medicine of Signal Transduction. Raven Press, New York (1990).

    Google Scholar 

  7. Cohen PTW, Brewis ND, Hughes V, Mann DJ: Protein serine/threonine phosphatases: an expanding family. FEBS Lett 268: 355–359 (1990).

    Article  PubMed  Google Scholar 

  8. Cormier P, Osborne HB, Bassez T, Bellé R, Mulner-Lorillon O: Protein phosphatase 2A from Xenopus oocytes: Characterization during meiotic cell division. FEBS Lett 295: 185–188 (1991).

    Article  PubMed  Google Scholar 

  9. Dean L, Elzen B, Tamaki S, Dunsmuir P, Bedbrook J: Differential expression of the eight genes of the petunia ribulose bisphosphate carboxylase small subunit multigene family. EMBO J 5: 3055–3061 (1985).

    Google Scholar 

  10. Dellaporta SL, Wood J, Hicks JB: Maize DNA miniprep. In. Molecular Biology of Plants: A Laboratory Manual, pp. 36–37. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York (1984).

    Google Scholar 

  11. Erondu NE, Donelson JE: Characterization of trypanosome protein phosphatase 1 and 2A catalytic subunits. Mol Biochem Parasitol 49: 303–314 (1991).

    Article  PubMed  Google Scholar 

  12. Hunter T: A thousand and one protein kinases. Cell 50: 823–829 (1987).

    Article  PubMed  Google Scholar 

  13. Ingebritsen TS, Cohen P: Protein phosphatases: Properties and role in cellular regulation. Science 221: 331–338 (1983).

    PubMed  Google Scholar 

  14. Jagiello I, Donella-Deana A, Szczegielniak J, Pinna LA, Muszyńska G: Identification of protein phosphatase activities in maize seedlings. Biochem Biophys Acta 1134: 129–136 (1992).

    Article  PubMed  Google Scholar 

  15. Khew-Goodall Y, Hemmings BA: Tissue-specific expression of mRNAs encoding α and β-catalytic subunits of protein phosphatase 2A. FEBS Lett 238: 265–268 (1988).

    Article  PubMed  Google Scholar 

  16. Khew-Goodall Y, Mayer RE, Maurer F, Stone SR, Hemmings BA. Structure and transcriptional regulation of protein phosphatase 2A catalytic subunit genes. Biochemistry 30: 89–97 (1991).

    PubMed  Google Scholar 

  17. Kinoshita N, Ohkura H, Yanagida M: Distinct, essential roles of type 1 and 2A protein phosphatases in the control of the fission yeast cell division cycle. Cell 63: 405–415 (1990).

    Article  PubMed  Google Scholar 

  18. Kitagawa T, Tahira T, Ikeda I, Kikuchi K, Tsuiki S, Sigimura T, Nagao M: Molecular cloning of cDNA for the catalytic subunit of rat liver type 2A protein phosphatase, and detection of high levels of expression of the gene in normal and cancer cells. Biochim Biophys Acta 951: 123–129 (1988).

    PubMed  Google Scholar 

  19. Lin PP-C, Mori T, Key JL: Phosphoprotein phosphatase of soybean hypocotyls. Plant Physiol. 66: 368–374 (1980).

    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).

    PubMed  Google Scholar 

  21. Mackintosh C, Cohen P: Identification of high levels of type 1 and type 2A protein phosphatases in higher plants. Biochem J 262: 335–339 (1989).

    PubMed  Google Scholar 

  22. Mackintosh C, Coggins J, Cohen P: Subcellular distribution, detection of protein phosphatase 2C and identification of protein phosphatase 2A as the major quinate dehydrogenase phosphatase. Biochem J. 273: 733–738 (1990).

    Google Scholar 

  23. Mackintosh RW, Haycox G, Hardie DG, Cohen PTW: Identification by molecular cloning of two cDNA sequences from the plant Brassica napus which are very similar to mammalian protein phosphatase-1 and-2A. FEBS Lett 276: 156–160 (1990).

    Article  PubMed  Google Scholar 

  24. Nitschke K, Fleig U, Schell J, Palme K: Complementation of the cs dis2–11 cell cycle mutant of Schizosaccharomyces pombe by a protein phosphatase 1. EMBO J 11: 1327–1333 (1992).

    PubMed  Google Scholar 

  25. Orgad S, Brewis ND, Alphey L, Axton JM, Dudai Y, Cohen PTW: The structure of protein phosphatase 2A is as highly conserved as that of protein phosphatase 1. FEBS Lett 275: 44–48 (1990).

    Article  PubMed  Google Scholar 

  26. Polya GM, Haritou M: Purification and characterization of two wheat-embryo protein phosphatases. Bicchem J 251: 357–363 (1988).

    Google Scholar 

  27. Ronne H, Carlberg M, Hu GZ, Nehlin JO: Protein phosphatase 2A in Saccharomyces cerevisiae: Effects on cell growth and bud morphogenesis. Mol Cell Biol 11: 4876–4884 (1991).

    PubMed  Google Scholar 

  28. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    PubMed  Google Scholar 

  29. Siegl G, Mackintosh C, Stitt M: Sucrose-phosphate synthase is dephosphorylated by protein phosphatase 2A in spinach leaves. FEBS Lett 270: 198–202 (1990).

    Article  PubMed  Google Scholar 

  30. Smith RD, Walker JC: Isolation and expression of a maize type 1 protein phosphatase. Plant Physiol 97: 677–683 (1991).

    Google Scholar 

  31. Sneddon AA, Cohen PTW, Stark MJR: Saccharomyces cerevisiae protein phosphatase 2A performs an essential cellular function and is encoded by two genes. EMBO J 9: 4339–4346 (1990).

    PubMed  Google Scholar 

  32. Swofford DL: Phylogenetic Analysis Using Parsimony, Version 3.0, Computer program distributed by the Illinois Natural History Survey, Champaign, Illinois (1991)

  33. Trewavas A, Gilroy S: Signal transduction in plant cells. Trends in Genet 7: 356–361 (1991).

    Google Scholar 

  34. Wadzinsky BE, Heasley LE, Johnson GL: Multiplicity of protein serine-threonine phosphatases in PC12 pheocromocytoma and FTO-2B hepatoma cells. J Biol Chem 265: 21504–21508 (1990).

    PubMed  Google Scholar 

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Ariño, J., Pérez-Callejón, E., Cunillera, N. et al. Protein phosphatases in higher plants: multiplicity of type 2A phosphatases in Arabidopsis thaliana . Plant Mol Biol 21, 475–485 (1993). https://doi.org/10.1007/BF00028805

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