Plant Molecular Biology

, Volume 20, Issue 3, pp 367–375 | Cite as

Molecular characterization of type 1 serine/threonine phosphatases from Brassica oleracea

  • Sabine J. Rundle
  • June B. Nasrallah
Research Article
Received:
Accepted:

Abstract

We describe the isolation of cDNA clones encoding type 1 serine/threonine protein phosphatase (PP1) from Brassica oleracea stigmas. We demonstrate that PP1 form a multigene family in Brassica. Within their most conserved domain, these phosphatases are 80–90% identical at the amino acid level. One cDNA (BoPP1) was found to encode a protein that shows 78–80% sequence identity to maize, rabbit, and yeast PP1. The accumulation of BoPP1 mRNA is developmentally regulated. Varying levels of BoPP1-homologous transcripts were detected in leaves, cotyledons, pistils, anthers and roots. In addition, distinct species of BoPP1 transcripts accumulated at different stages of Brassica microspore development, and mature trinucleate microspores contained a unique BoPP1 mRNA species not found at other stages of the plant's life cycle. Lastly, we show by genomic Southern blots that the Brassica genome might contain homologues of the mammalian PP1 inhibitor-1.

Key words

Brassica oleracea inhibitor 1 microspore serine/threonine protein phosphatase 
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Arndt K, Styles C, Fink G: A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases. Cell 56: 527–537 (1989).Google Scholar
  2. 2.
    Axton J, Dombradi V, Cohen PT, Glover D: One of the protein phosphatase 1 isoenzymes in Drosophila is essential for mitosis. Cell 63: 33–46 (1990).Google Scholar
  3. 3.
    Berndt N, Campbell D, Caudwell B, Cohen P, da Cruz e Silva E, da Cruz e Silva O, Cohen PT: Isolation and sequence analysis of a cDNA clone encoding a type-1 protein phosphatase catalytic subunit: homology with protein phosphatase 2A. FEBS Lett 232: 340–346 (1987).Google Scholar
  4. 4.
    Bingham P, Lewis R, Rubin G: Cloning of DNA sequences from the white locus of D. melanogaster by a novel and general method. Cell 25: 693–704 (1981).Google Scholar
  5. 5.
    Booker R, Beach D: Involvement of a type 1 protein phosphatase encoded by bws1 + in fission yeast mitotic control. Cell 57: 1009–1016 (1989).Google Scholar
  6. 6.
    Carter P, Nimmo H, Fewson C, Wilkins M: Bryophyllum fedtschenkoi protein phosphatase type 2A dephosphorylates phosphoenolpyruvate carboxylase. FEBS Lett 263: 233–236 (1990).Google Scholar
  7. 7.
    Cohen P: The structure and regulation of protein phosphatases. Annu Rev Biochem 58: 453–508 (1989).Google Scholar
  8. 8.
    Cohen PT: Two isoforms of protein phosphatase 1 may be produced from the same gene. FEBS Lett 232: 17–23 (1988).Google Scholar
  9. 9.
    Cohen P, Cohen PT: Protein phosphatases come of age. J Biol Chem 264: 21435–21438 (1989).Google Scholar
  10. 10.
    Dombradi V, Axton J, Brewis N, da Cruz e Silva E, Alphey L, Cohen PT: Drosophila contains three genes that encode distinct isoforms of protein phosphatase 1. Eur J Biochem 194: 739–745 (1990).Google Scholar
  11. 11.
    Dombradi V, Axton J, Glover D, Cohen PT: Cloning and chromosomal localization of Drosophila cDNA encoding the catalytic subunit of protein phosphatase 1-α. Eur J Biochem 183: 603–610 (1989).Google Scholar
  12. 12.
    Doonan J, Morris R: The bimG gene of Aspergillus nidulans, required for completion of anaphase, encodes a homolog of mammalian phosphoprotein phosphatase 1. Cell 57: 987–996 (1989).Google Scholar
  13. 13.
    Elbrecht A, DiRenzo J, Smith R, Shenolikar S: Molecular cloning of protein phosphatase inhibitor-1 and its expression in rat and rabbit tissues. J Biol Chem 265: 13415–13418 (1990).Google Scholar
  14. 14.
    Feinberg A, Vogelstein B: A technique for radiolabeling DNA restriction fragments to a high specific activity. Anal Biochem 132: 6–13 (1983).Google Scholar
  15. 15.
    Guerini D, Klee C: Cloning of human calcineurin A: evidence for two isozymes and identification of a polyproline structural domain. Proc Natl Acad Sci USA 86: 9183–9187 (1989).Google Scholar
  16. 16.
    Henikoff S: A unidirectional with Exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28: 351–359 (1984).Google Scholar
  17. 17.
    Hinnebusch A: Evidence for translational regulation of the activator of general amino acid control in yeast. Proc Natl Acad Sci USA 81: 6442–6446 (1984).Google Scholar
  18. 18.
    Hubbard M, Cohen P: Regulation of protein phosphatase-1G from rabbit skeletal muscle. Eur J Biochem 1186: 701–709 (1989).Google Scholar
  19. 19.
    Huber S, Huber J: Activation of sucrose-phosphate synthase for darkened spinach leaves by an endogenous protein phosphatase. Arch Biochem Biophys 282: 4421–4426 (1990).Google Scholar
  20. 20.
    Khew-Goodall Y, Hemmings B: Tissue-specific expression of mRNAs encoding α and β catalytic subunits of protein phosphatase 2A. FEBS Lett 238: 265–268 (1988).Google Scholar
  21. 21.
    MacKintosh C, Coggins J, Cohen P: Plant protein phosphatases. Biochem J 273: 733–738 (1991).Google Scholar
  22. 22.
    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).Google Scholar
  23. 23.
    MacKintosh R, Haycox G, Hardie D, Cohen PT: Identification by molecular cloning of two cDNA sequences from the plant Brassica napus which are very similar to mammalian protein phosphatases 1 and 2A. FEBS Lett 276: 156–160 (1990).Google Scholar
  24. 24.
    Maniatis T, Fritsch E, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).Google Scholar
  25. 25.
    Nasrallah J, Yu S, Nasrallah M: Self-incompatibility genes of Brassica oleracea: expression, isolation and structure. Proc Natl Acad Sci USA 85: 5551–5555 (1988).Google Scholar
  26. 26.
    Ohkura H, Kinoshita N, Miyatani S, Toda T, Yanagida M: The fission yeast dis2 + gene required for chromosome disjoining encodes one of two putative type 1 protein phosphatases. Cell 57: 997–1007 (1989).Google Scholar
  27. 27.
    Pearson W, Lipman D: Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85: 2444–2448 (1988).Google Scholar
  28. 28.
    Ruberti I, Sessa G, Lucchetti S, Morelli G: A novel class of plant proteins containing a homeodomain with a closely linked leucine zipper motif. EMBO J 10: 1787–1791 (1991).Google Scholar
  29. 29.
    Rundle S, Zielinski R: Organization and expression of two tandemly oriented genes encoding ribulosebisphosphate carboxylase/oxygenase activase in barley. J Biol Chem 266: 4677–4685 (1991).Google Scholar
  30. 30.
    Sato T, Thorsness M, Kandasamy M, Nishio T, Hirai M, Nasrallah J, Nasrallah M: Activity of an S locus gene promoter in pistils and anthers of transgenic Brassica. Plant Cell 3: 867–876 (1991).Google Scholar
  31. 31.
    Siegl G, MacKintosh C, Stitt M: Sucrose-phosphate synthase is dephosphorylated by protein phosphatase 2A in spinach leaves. FEBS Lett 270: 198–202 (1990).Google Scholar
  32. 32.
    Smith R, Walker J: Isolation and expression of a maize type 1 protein phosphatase. Plant Physiol 97: 677–683 (1991).Google Scholar
  33. 33.
    Stein J, Howlett B, Boyes D, Nasrallah M, Nasrallah J: Molecular cloning of a putative receptor protein kinase gene encoded at the self-incompatibility locus of Brassica oleracea. Proc Natl Acad Sci USA 88: 8816–8820 (1991).Google Scholar
  34. 34.
    Tonks N: Protein phosphatases: key players in the regulation of cell function. Curr Opin Cell Biol 2: 1114–1124 (1990).Google Scholar

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Sabine J. Rundle
    • 1
  • June B. Nasrallah
    • 1
  1. 1.Section of Plant Biology, Division of Biological SciencesCornell UniversityIthacaUSA

Personalised recommendations