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Molecular Methods for the Study of Signal Transduction in Plants

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Cyclic Nucleotide Signaling in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1016))

Abstract

Novel and improved analytical methods have led to a rapid increase in our understanding of the molecular mechanism underlying plant signal transduction. Progress has been made both at the level of single-component analysis and in vivo imaging as well as at the systems level where transcriptomics and particularly phosphoproteomics afford a window into complex biological responses. Here we review the role of the cyclic nucleotides cAMP and cGMP in plant signal transduction as well as the discovery and biochemical and biological characterization of an increasing number of complex multi-domain nucleotide cyclases that catalyze the synthesis of cAMP and cGMP from ATP and GTP, respectively.

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References

  1. Stael S, Wursinger B, Mair A, Mehlmer N, Vothknecht UC, Teige M (2012) Plant organellar calcium signalling: an emerging field. J Exp Bot 63:1525–1542

    Article  PubMed  CAS  Google Scholar 

  2. Hirschi KD (2004) The calcium conundrum. Both versatile nutrient and specific signal. Plant Physiol 136:2438–2442

    Article  PubMed  CAS  Google Scholar 

  3. Sanders D, Pelloux J, Brownlee C, Harper JF (2002) Calcium at the crossroads of signaling. Plant Cell 14(Suppl):S401–S417

    PubMed  CAS  Google Scholar 

  4. Kudla J, Batistic O, Hashimoto K (2010) Calcium signals: the lead currency of plant information processing. Plant Cell 22:541–563

    Article  PubMed  CAS  Google Scholar 

  5. Reid RJ, Smith FA (1992) Measurement of calcium fluxes in plants using 45Ca. Planta 186:558–566

    Article  CAS  Google Scholar 

  6. Felle HH, Kondorosi E, Kondorosi A, Schultze M (1999) Elevation of the cytosolic free [Ca2+] is indispensible for the transduction of the nod factor signal in alfalfa. Plant Physiol 121:273–279

    Article  PubMed  CAS  Google Scholar 

  7. Felle H (1988) Auxin causes oscillations of cytosolic free calcium and pH in Zea mays coleoptiles. Planta 174:495–499

    Article  CAS  Google Scholar 

  8. Newman IA (2001) Ion transport in roots: measurement of fluxes using ion-selective microelectrodes to characterize transporter function. Plant Cell Environ 24:1–14

    Article  PubMed  CAS  Google Scholar 

  9. Gilliham M, Sullivan W, Tester M, Tyerman SD (2006) Simultaneous flux and current measurement from single plant protoplasts reveals a strong link between K+ fluxes and current, but no link between Ca2+ fluxes and current. Plant J 46:134–144

    Article  PubMed  CAS  Google Scholar 

  10. Donaldson L, Ludidi N, Knight MR, Gehring C, Denby K (2004) Salt and osmotic stress cause rapid increases in Arabidopsis thaliana cGMP levels. FEBS Lett 569:317–320

    Article  PubMed  CAS  Google Scholar 

  11. Knight MR, Campbell AK, Smith SM, Trewavas AJ (1991) Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. Nature 352:524–526

    Article  PubMed  CAS  Google Scholar 

  12. Mehlmer N, Parvin N, Hurst CH, Knight MR, Teige M, Vothknecht UC (2012) A toolset of aequorin expression vectors for in planta ­studies of subcellular calcium concnetration in Arabidopsis thaliana. J Exp Bot 63:1751–1761

    Article  PubMed  CAS  Google Scholar 

  13. Okumoto S (2012) Quantitative imaging using genetically encoded sensors for small molecules in plants. Plant J 70:108–117

    Article  PubMed  CAS  Google Scholar 

  14. Choi WG, Swanson SJ, Gilroy S (2012) High-resolution imaging of Ca2+, redox status, ROS and pH using GFP biosensors. Plant J 70:118–128

    Article  PubMed  CAS  Google Scholar 

  15. Souslova EA, Chudakov DM (2007) Genetically encoded intracellular sensors based on fluorescent proteins. Biochemstry (Mosc) 72:683–697

    Article  CAS  Google Scholar 

  16. Allen GJ, Kwak JM, Chu SP, Llopas J, Tsien RY, Harper JF et al (1999) Cameleon calcium indicator reports cytoplasmic calcium dynamics in Arabidopsis guard cells. Plant J 19:735–747

    Article  PubMed  CAS  Google Scholar 

  17. Krebs M, Held K, Binder A, Hashimoto K, Den Herder G, Parniske M et al (2012) FRET-based genetically encoded sensors allow high-resolution live cell imaging of Ca2+ dynamics. Plant J 69:181–192

    Article  CAS  Google Scholar 

  18. Gehring C (2010) Adenyl cyclases and cAMP in plant signaling—past and present. Cell Commun Signal 8:15

    Article  PubMed  Google Scholar 

  19. Li WW, Luan S, Schreiber SL, Assmann SM (1994) Cyclic-AMP stimulates K+ channel activity in mesophyll-cells of Vicia faba. Plant Physiol 106:957–961

    Article  PubMed  CAS  Google Scholar 

  20. Lemtiri-Chlieh F, Berkowitz GA (2004) Cyclic adenosine monophosphate regulates calcium channels in the plasma membrane of Arabidopsis leaf guard and mesophyll cells. J Biol Chem 279:35306–35312

    Article  PubMed  CAS  Google Scholar 

  21. Leng Q, Mercier RW, Yao W, Berkowitz GA (1999) Cloning and first functional characterization of a plant cyclic nucleotide-gated cation channel. Plant Physiol 121:753–761

    Article  PubMed  CAS  Google Scholar 

  22. Zelman AK, Dawe A, Gehring C, Berkowitz GA (2012) Evolutionary and structural perspectives of plant cyclic nucleotide-gated cation channels. Front Plant Sci 3:95

    Article  PubMed  Google Scholar 

  23. Ali R, Ma W, Lemtiri-Chlieh F, Tsaltas D, Leng Q, von Bodman S et al (2007) Death don’t have no mercy and neither does calcium: Arabidopsis CYCLIC NUCLEOTIDE GATED CHANNEL2 and innate immunity. Plant Cell 19:1081–1095

    Article  PubMed  CAS  Google Scholar 

  24. Maathuis FJ, Sanders D (2001) Sodium uptake in Arabidopsis roots is regulated by cyclic nucleotides. Plant Physiol 127:1617–1625

    Article  PubMed  CAS  Google Scholar 

  25. Maathuis FM (2006) cGMP modulates gene transcription and cation transport in Arabidopsis roots. Plant J 45:700–711

    Article  PubMed  CAS  Google Scholar 

  26. Whiteman SA, Nuhse TS, Ashford DA, Sanders D, Maathuis FJ (2008) A proteomic and phosphoproteomic analysis of Oryza sativa plasma membrane and vacuolar membrane. Plant J 56:146–156

    Article  PubMed  CAS  Google Scholar 

  27. Isner JC, Nuhse T, Maathuis FJ (2012) The cyclic nucleotide cGMP is involved in plant hormone signalling and alters phosphorylation of Arabidopsis thaliana root proteins. J Exp Bot 63:3199–3205

    Article  PubMed  CAS  Google Scholar 

  28. Ludidi N, Gehring C (2003) Identification of a novel protein with guanylyl cyclase activity in Arabidopsis thaliana. J Biol Chem 278:6490–6494

    Article  PubMed  CAS  Google Scholar 

  29. Meier S, Ruzvidzo O, Morse M, Donaldson L, Kwezi L, Gehring C (2010) The Arabidopsis wall associated kinase-like 10 gene encodes a functional guanylyl cyclase and is co-expressed with pathogen defense related genes. PLoS One 5:e8904

    Article  PubMed  Google Scholar 

  30. Kwezi L, Meier S, Mungur L, Ruzvidzo O, Irving H, Gehring C (2007) The Arabidopsis thaliana brassinosteroid receptor (AtBRI1) contains a domain that functions as a guanylyl cyclase in vitro. PLoS One 2:e449

    Article  PubMed  Google Scholar 

  31. Qi Z, Verma R, Gehring C, Yamaguchi Y, Zhao Y, Ryan CA et al (2010) Ca2+ signaling by plant Arabidopsis thaliana Pep peptides depends on AtPepR1, a receptor with guanylyl cyclase activity, and cGMP-activated Ca2+ ­channels. Proc Natl Acad Sci U S A 107:21193–21198

    Article  PubMed  CAS  Google Scholar 

  32. Kwezi L, Ruzvidzo O, Wheeler JI, Govender K, Iacuone S, Thompson PE et al (2011) The phytosulfokine (PSK) receptor is capable of guanylate cyclase activity and enabling cyclic GMP-dependent signaling in plants. J Biol Chem 286:22580–22588

    Article  PubMed  CAS  Google Scholar 

  33. Mulaudzi T, Ludidi N, Ruzvidzo O, Morse M, Hendricks N, Iwuoha E et al (2011) Identification of a novel Arabidopsis thaliana nitric oxide-binding molecule with guanylate cyclase activity in vitro. FEBS Lett 585:2693–2697

    Article  PubMed  CAS  Google Scholar 

  34. Irving HR, Kwezi L, Wheeler JI, Gehring C (2012) Moonlighting kinases with guanylate cyclase activity can tune regulatory signal networks. Plant Signal Behav 7:201–204

    Article  PubMed  CAS  Google Scholar 

  35. Moutinho A, Hussey PJ, Trewavas AJ, Malho R (2001) cAMP acts as a second messenger in pollen tube growth and reorientation. Proc Natl Acad Sci U S A 98:10481–10486

    Article  PubMed  CAS  Google Scholar 

  36. DeYoung BJ, Innes RW (2006) Plant NBS-LRR proteins in pathogen sensing and host defense. Nat Immunol 7:1243–1249

    Article  PubMed  CAS  Google Scholar 

  37. Gehring CA, Irving HR (2003) Natriuretic peptides—a class of heterologous molecules in plants. Int J Biochem Cell Biol 35:1318–1322

    Article  PubMed  CAS  Google Scholar 

  38. Meier S, Madeo L, Ederli L, Donaldson L, Pasqualini S, Gehring C (2009) Deciphering cGMP signatures and cGMP-dependent pathways in plant defence. Plant Signal Behav 4:307–309

    Article  PubMed  CAS  Google Scholar 

  39. Clouse SD (2011) Brassinosteroid signal transduction: from receptor kinase activation to transcriptional networks regulating plant development. Plant Cell 23:1219–1230

    Article  PubMed  CAS  Google Scholar 

  40. Durner J, Klessig DF (1999) Nitric oxide as a signal in plants. Curr Opin Plant Biol 2:369–374

    Article  PubMed  CAS  Google Scholar 

  41. Durner J, Wendehenne D, Klessig DF (1998) Defense gene induction in tobacco by nitric oxide, cyclic GMP, and cyclic ADP-ribose. Proc Natl Acad Sci U S A 95:10328–10333

    Article  PubMed  CAS  Google Scholar 

  42. Clarke A, Desikan R, Hurst RD, Hancock JT, Neill SJ (2000) NO way back: nitric oxide and programmed cell death in Arabidopsis thaliana suspension cultures. Plant J 24:667–677

    Article  PubMed  CAS  Google Scholar 

  43. Pasqualini S, Meier S, Gehring C, Madeo L, Fornaciari M, Romano B et al (2009) Ozone and nitric oxide induce cGMP-dependent and -independent transcription of defence genes in tobacco. New Phytol 181:860–870

    Article  PubMed  CAS  Google Scholar 

  44. Boon EM, Huang SH, Marletta MA (2005) A molecular basis for NO selectivity in soluble guanylate cyclase. Nat Chem Biol 1:53–59

    Article  PubMed  CAS  Google Scholar 

  45. Isner JC, Maathuis FJ (2011) Measurement of cellular cGMP in plant cells and tissues using the endogenous fluorescent reporter FlincG. Plant J 65:329–334

    Article  PubMed  CAS  Google Scholar 

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Irving, H.R., Gehring, C. (2013). Molecular Methods for the Study of Signal Transduction in Plants. In: Gehring, C. (eds) Cyclic Nucleotide Signaling in Plants. Methods in Molecular Biology, vol 1016. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-441-8_1

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  • DOI: https://doi.org/10.1007/978-1-62703-441-8_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-440-1

  • Online ISBN: 978-1-62703-441-8

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