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Expression analysis and subcellular localization of the Arabidopsis thaliana G-protein β-subunit AGB1

  • Cell Biology and Morphogenesis
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Abstract

Heterotrimeric GTP-binding proteins (G-proteins), consisting of Gα, Gβ, and Gγ subunits, function as molecular switches in many eukaryotic signal transduction pathways. In contrast to many eukaryotes, plants contain very few G-protein subunit isoforms that mediate a diverse array of signalling functions. We investigated the contribution of cell type-specific expression and subcellular localization to this multifunctional signalling capacity for the Arabidopsis thaliana Gβ subunit, AGB1. Analysis of AGB1 promoter::β-glucuronidase (GUS) fusions in germinating seeds, seedlings, and flowering plants revealed that AGB1 is widely expressed throughout development in a complex manner. As well as demonstrating similarities to existing Arabidopsis G-protein subunit expression data, several features of the AGB1 expression pattern align closely with known or proposed G-protein functions. A C-terminal AGB1-green fluorescent protein (GFP) fusion was localized at the plasma membrane and in the nucleus of leaf epidermal cells, trichomes and root cells, supporting previous evidence that plant G-protein functionality relies on subcellular compartmentalization.

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Abbreviations

CLSM:

Confocal laser scanning microscopy

ER:

Endoplasmic reticulum

GFP:

Green fluorescent protein

G-protein:

Heterotrimeric GTP-binding protein

GPCR:

G-protein-coupled receptor

GUS:

β-glucuronidase

rRNA:

ribosomal RNA

References

  • Assmann SM (2005) G proteins go green: a plant G protein signaling FAQ sheet. Science 310:71–73

    Article  PubMed  CAS  Google Scholar 

  • Bechtold N, Ellis J, Pelletier G (1993) In planta Agrobacterium-mediated gene transfer of adult Arabidopsis thaliana plants. C R Seances Soc Biol Fil 316:1194–1199

    CAS  Google Scholar 

  • Casey PJ (1995) Mechanisms of protein prenylation and role in G-protein function. Biochem Soc Trans 23:161–166

    PubMed  CAS  Google Scholar 

  • Chen JG, Gao YJ, Jones AM (2006a) Differential roles of Arabidopsis heterotrimeric G-protein subunits in modulating cell division in roots. Plant Physiol 141:887–897

    Article  PubMed  CAS  Google Scholar 

  • Chen ZY, Hartmann HA, Wu MJ, Friedman EJ, Chen JG, Pulley M, Schulze-Lefert P, Panstruga R, Jones AM (2006b) Expression analysis of the AtMLO gene family encoding plant-specific seven-transmembrane domain proteins. Plant Mol Biol 60:583–597

    Article  PubMed  CAS  Google Scholar 

  • Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range cloning system for gram negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351

    Article  PubMed  CAS  Google Scholar 

  • Etheridge N, Trusov Y, Verbelen JP, Botella JR (1999) Characterization of ATDRG1, a member of a new class of GTP-binding proteins in plants. Plant Mol Biol 39:1113–1126

    Article  PubMed  CAS  Google Scholar 

  • Gerlach WL, Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res 7:1869–1885

    Article  PubMed  CAS  Google Scholar 

  • Gotta M, Ahringer J (2001) Distinct roles for G alpha and G beta gamma in regulating spindle position and orientation in Caenorhabditis elegans embryos. Nat Cell Biol 3:297–300

    Article  PubMed  CAS  Google Scholar 

  • Gutierrez-Alcala G, Gotor C, Meyer AJ, Fricker M, Vega JM, Romero LC (2000) Glutathione biosynthesis in Arabidopsis trichome cells. Proc Natl Acad Sci USA 97:11108–11113

    Article  PubMed  CAS  Google Scholar 

  • Hampoelz B, Knoblich JA (2004) Heterotrimeric G proteins: new tricks for an old dog. Cell 119:453–456

    Article  PubMed  CAS  Google Scholar 

  • Haseloff J (1999) GFP variants for multispectral imaging of living cells. In: Sullivan KF, Kay SA (eds) Methods in cell biology: green fluorescent proteins. Academic, San Diego, pp 139–151

    Google Scholar 

  • Haseloff J, Siemering KR (1998) The uses of green fluorescent protein in plants. In: Chalfie M, Kain S (eds) Green fluorescent protein: properties, applications, and protocols. Wiley-Liss, New York, pp 191–220

    Google Scholar 

  • Haseloff J, Siemering KR, Prasher DC, Hodge S (1997) Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc Natl Acad Sci USA 94:2122–2127

    Article  PubMed  CAS  Google Scholar 

  • Hoekema A, Hirsch PR, Hookyaas PJJ, Schilperoort RA (1983) A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti plasmid. Nature 303:179–180

    Article  CAS  Google Scholar 

  • Huang H, Weiss CA, Ma H (1994) Regulated expression of the Arabidopsis G protein α subunit gene GPA1. Int J Plant Sci 155:3–14

    Article  CAS  Google Scholar 

  • Humphrey TV, Botella JR (2001) Re-evaluation of the cytokinin receptor role of the Arabidopsis gene GCR1. J Plant Physiol 158:645–653

    Article  CAS  Google Scholar 

  • Jiang K, Zhang SB, Lee S, Tsai G, Kim K, Huang HY, Chilcott C, Zhu T, Feldman LJ (2006) Transcription profile analyses identify genes and pathways central to root cap functions in maize. Plant Mol Biol 60:343–363

    Article  PubMed  CAS  Google Scholar 

  • Jones AM, Assmann SM (2004) Plants: the latest model system for G-protein research. EMBO Rep 5:572–578

    Article  PubMed  CAS  Google Scholar 

  • Kaydamov C, Tewes A, Adler K, Manteuffel R (2000) Molecular characterization of cDNAs encoding G protein α and β subunits and study of their temporal and spatial expression patterns in Nicotiana plumbaginifolia Viv. Biochim Biophys Acta 25:1–3

    Google Scholar 

  • Kohler RH (1998) GFP for in vivo imaging of subcellular structures in plant cells. Trends Plant Sci 3:317–320

    Article  Google Scholar 

  • Ma H, Yanofsky MF, Meyerowitz EM (1990) Molecular cloning and characterization of GPA1, a G protein α-subunit gene from Arabidopsis thaliana. Proc Natl Acad Sci USA 87:3821–3825

    Article  PubMed  CAS  Google Scholar 

  • Malbon CC (2005) G proteins in development. Nat Rev Mol Cell Biol 6:689–701

    Article  PubMed  CAS  Google Scholar 

  • Mason MG, Botella JR (2000) Completing the heterotrimer: isolation and characterization of an Arabidopsis thaliana G protein γ-subunit cDNA. Proc Natl Acad Sci USA 97:14784–14788

    Article  PubMed  CAS  Google Scholar 

  • Mason MG, Botella JR (2001) Isolation of a novel G-protein γ-subunit from Arabidopsis thaliana and its interaction with Gβ. Biochim Biophys Acta 1520:147–153

    PubMed  CAS  Google Scholar 

  • Matkin OA, Chandler PA (1979) The U. C.–type soil mixes. In: Baker KF (ed) The UC system for producing healthy container-grown plants. University of California, Division of Agricultural Sciences, Agricultural Experiment Station, Extension Service, Berkeley, pp 68–85

  • Mishra G, Zhang WH, Deng F, Zhao J, Wang XM (2006) A bifurcating pathway directs abscisic acid effects on stomatal closure and opening in Arabidopsis. Science 312:264–266

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Mylne JS, Botella JR (1998) Binary vectors for sense and antisense expression of Arabidopsis ESTs. Plant Mol Biol Rep 16:257–262

    Article  CAS  Google Scholar 

  • Obrdlik P, Neuhaus G, Merkle T (2000) Plant heterotrimeric G protein beta subunit is associated with membranes via protein interactions involving coiled-coil formation. FEBS Lett 476:208–212

    Article  PubMed  CAS  Google Scholar 

  • Offermanns S (2003) G-proteins as transducers in transmembrane signalling. Prog Biophys Mol Biol 83:101–130

    Article  PubMed  CAS  Google Scholar 

  • Ohta S, Mita S, Hattori T, Nakamura K, Benfey PN, Ren L, Chua NH (1990) Construction and expression in tobacco of a beta-glucuronidase (GUS) reporter gene containing an intron within the coding sequence: tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development. Plant Cell Physiol 31:805–814

    CAS  Google Scholar 

  • Perfus-Barbeoch L, Jones AM, Assmann SM (2004) Plant heterotrimeric G protein function: insights from Arabidopsis and rice mutants. Curr Opin Plant Biol 7:719–731

    Article  PubMed  CAS  Google Scholar 

  • Peškan T, Oelmüller R (2000) Heterotrimeric G-protein beta-subunit is localized in the plasma membrane and nuclei of tobacco leaves. Plant Mol Biol 42:915–922

    Article  PubMed  Google Scholar 

  • Pierce KL, Premont RT, Lefkowitz RJ (2002) Seven-transmembrane receptors. Nat Rev Mol Cell Biol 3:639–650

    Article  PubMed  CAS  Google Scholar 

  • Rasband W (2005) Image J 1.33u. In. National Institutes of Health, USA

  • Rietz S, Holk A, Scherer GFE (2004) Expression of the patatin-related phospholipase A gene AtPLA IIA in Arabidopsis thaliana is up-regulated by salicylic acid, wounding, ethylene, and iron and phosphate deficiency. Planta 219:743–753

    Article  PubMed  CAS  Google Scholar 

  • Rolland F, Baena-Gonzalez E, Sheen J (2006) Sugar sensing and signaling in plants: conserved and novel mechanisms. Annu Rev Plant Biol 57:675–709

    Article  PubMed  CAS  Google Scholar 

  • Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6:1653–1663

    Article  PubMed  CAS  Google Scholar 

  • Simonds WF, Woodard GE, Zhang JH (2004) Assays of nuclear localization of R7/G beta(5) complexes. In: Regulators of G-protein signaling, Pt B, pp 210–223

  • Svab Z, Hajdukiewicz P, Maliga P (1995) Generation of transgenic tobacco plants by cocultivation of leaf disks with Agrobacterium pPZP binary vectors. In: Maliga P, Klessig DF, Cashmore AR, Gruissem W, Varner JE (eds) Methods in plant molecular biology: a laboratory course manual. Cold Spring Harbor Laboratory Press, Plainview, pp 55–77

    Google Scholar 

  • Takahashi N, Kuroda H, Kuromori T, Hirayama T, Seki M, Shinozaki K, Shimada H, Matsui M (2004) Expression and interaction analysis of Arabidopsis Skp1-related genes. Plant Cell Physiol 45:83–91

    Article  PubMed  CAS  Google Scholar 

  • Traw MB, Bergelson J (2003) Interactive effects of jasmonic acid, salicylic acid, and gibberellin on induction of trichomes in Arabidopsis. Plant Physiol 133:1367–1375

    Article  PubMed  CAS  Google Scholar 

  • Trusov Y, Rookes JE, Chakravorty D, Armour D, Schenk PM, Botella JR (2006) Heterotrimeric G proteins facilitate Arabidopsis resistance to necrotrophic pathogens and are involved in jasmonate signaling. Plant Physiol 140:210–220

    Article  PubMed  CAS  Google Scholar 

  • Ullah H, Chen JG, Young JC, Im KH, Sussman MR, Jones AM (2001) Modulation of cell proliferation by heterotrimeric G protein in Arabidopsis. Science 292:2066–2069

    Article  PubMed  CAS  Google Scholar 

  • Ullah H, Chen JG, Wang SC, Jones AM (2002) Role of a heterotrimeric G protein in regulation of Arabidopsis seed germination. Plant Physiol 129:897–907

    Article  PubMed  CAS  Google Scholar 

  • Ullah H, Chen JG, Temple B, Boyes DC, Alonso JM, Davis KR, Ecker JR, Jones AM (2003) The beta-subunit of the Arabidopsis G protein negatively regulates auxin-induced cell division and affects multiple developmental processes. Plant Cell 15:393–409

    Article  PubMed  CAS  Google Scholar 

  • Viehweger K, Schwartze W, Schumann B, Lein W, Roos W (2006) The Gα protein controls a pH-dependent signal path to the induction of phytoalexin biosynthesis in Eschscholzia californica. Plant Cell 18:1510–1523

    Article  PubMed  CAS  Google Scholar 

  • Wang XQ, Ullah H, Jones AM, Assmann SM (2001) G protein regulation of ion channels and abscisic acid signaling in Arabidopsis guard cells. Science 292:2070–2072

    Article  PubMed  CAS  Google Scholar 

  • Weiss CA, Huang H, Ma H (1993) Immunolocalization of the G-protein α-subunit encoded by the GPA1 gene in Arabidopsis. Plant Cell 5:1513–1528

    Article  PubMed  CAS  Google Scholar 

  • Weiss CA, Garnaat CW, Mukai K, Hu Y, Ma H (1994) Isolation of cDNAs encoding guanine nucleotide-binding protein β-subunit homologues from maize (ZGB1) and Arabidopsis (AGB1). Proc Natl Acad Sci USA 91:9554–9558

    Article  PubMed  CAS  Google Scholar 

  • Weiss CA, White E, Huang H, Ma H (1997) The G protein α-subunit (GPα1) is associated with the ER and the plasma membrane in meristematic cells of Arabidopsis and cauliflower. FEBS Lett 407:361–367

    Article  PubMed  CAS  Google Scholar 

  • White MA, Anderson RGW (2005) Signaling networks in living cells. Annu Rev Pharmacol Toxicol 45:587–603

    Article  PubMed  CAS  Google Scholar 

  • Wienkoop S, Zoeller D, Ebert B, Simon-Rosin U, Fisahn J, Glinski M, Weckwerth W (2004) Cell-specific protein profiling in Arabidopsis thaliana trichomes: identification of trichome-located proteins involved in sulfur metabolism and detoxification. Phytochemistry 65:1641–1649

    Article  PubMed  CAS  Google Scholar 

  • Willard FS, Crouch MF (2000) Nuclear and cytoskeletal translocation and localization of heterotrimeric G-proteins. Immunol Cell Biol 78:387–394

    Article  PubMed  CAS  Google Scholar 

  • Zhao DZ, Ni WM, Feng BM, Han TF, Petrasek MG, Ma H (2003) Members of the Arabidopsis-SKP1-like gene family exhibit a variety of expression patterns and may play diverse roles in Arabidopsis. Plant Physiol 133:203–217

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This research was supported by an Australian Research Council Discovery grant (DP0344924). The authors thank the members of the Plant Genetic Engineering Laboratory for their helpful discussions and proofreading. We are grateful to Dr. T. V. Humphrey for the pBS-GFP construct, Dr. J. S. Mylne for the pAOV-intron-GUS and pSOV2 constructs, Dr. K. Siemering for the mGFP5-ER Arabidopsis seeds, and Dr. A. Gnanasambandam for the non-targeted mGFP5. D. J. A. was supported by an Australian Postgraduate Research Award from the Australian Government.

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  1. David J. Anderson

    Present address: The Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD, 4072, Australia

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  1. Plant Genetic Engineering Laboratory, School of Integrative Biology, The University of Queensland, St. Lucia, QLD, 4072, Australia

    David J. Anderson & José R. Botella

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  1. David J. Anderson
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Correspondence to José R. Botella.

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Communicated by P. Lakshmanan.

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Anderson, D.J., Botella, J.R. Expression analysis and subcellular localization of the Arabidopsis thaliana G-protein β-subunit AGB1. Plant Cell Rep 26, 1469–1480 (2007). https://doi.org/10.1007/s00299-007-0356-1

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