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Plant Molecular Biology

, Volume 65, Issue 5, pp 603–614 | Cite as

Distinct repressing modules on the distal region of the SBP2 promoter contribute to its vascular tissue-specific expression in different vegetative organs

  • Rejane L. Freitas
  • Claudine M. Carvalho
  • Luciano G. Fietto
  • Marcelo E. Loureiro
  • Andrea M. Almeida
  • Elizabeth P. B. Fontes
Article

Abstract

The Glycine max sucrose binding protein (GmSBP2) promoter directs vascular tissue-specific expression of reporter genes in transgenic tobacco. Here we showed that an SBP2-GFP fusion protein under the control of the GmSBP2 promoter accumulates in the vascular tissues of vegetative organs, which is consistent with the proposed involvement of SBP in sucrose transport-dependent physiological processes. Through gain-of-function experiments we confirmed that the tissue-specific determinants of the SBP2 promoter reside in the distal cis-regulatory domain A, CRD-A (position −2000 to −700) that is organized into a modular configuration to suppress promoter activity in tissues other than vascular tissues. The four analyzed CRD-A sub-modules, designates Frag II (−1785/−1508), Frag III (−1507/−1237), Frag IV (−1236/−971) and Frag V (−970/−700), act independently to alter the constitutive pattern of −92pSBP2-mediated GUS expression in different organs. Frag V fused to −92pSBP2-GUS restored the tissue-specific pattern of the full-length promoter in the shoot apex, but not in other organs. Likewise, Frag IV confined GUS expression to the vascular bundle of leaves, whereas Frag II mediated vascular specific expression in roots. Strong stem expression-repressing elements were located at positions −1485 to −1212, as Frag III limited GUS expression to the inner phloem. We have also mapped a procambium silencer to the consensus sequence CAGTTnCaAccACATTcCT which is located in both distal and proximal upstream modules. Fusion of either repressing element-containing module to the constitutive −92pSBP2 promoter suppresses GUS expression in the elongation zone of roots. Together our results demonstrate the unusual aspect of distal sequences negatively controlling tissue-specificity of a plant promoter.

Keywords

Transcriptional regulation Tissue-specific regulation Repressing elements Promoter distal regions Sucrose binding protein Cis-acting elements Vascular tissue-specific expression 

Abbreviations

GmSBP2

Glycine max sucrose-binding protein 2

VfSBPL

Vicia faba sucrose-binding protein-like protein

PCR

polymerase chain reaction

CRD

cis-regulatory domain

GUS

β-glucuronidase

Notes

Acknowledgements

We are grateful to Dr. Simon L. Elliot for critically reading the manuscript. This research was supported by the Brazilian Government Agencies CNPq grant 50.6119/2004-1 and 470878/2006-1 (to E.P.B.F.) and FAPEMIG grant EDT 560/05 and EDT 523/07 (to E.P.B.F.). R.L.F. was supported by a graduate fellowship from the Brazilian Government Agency CAPES. C.M.C. is a FAPEMIG postdoctoral fellow (CBB 00112/07).

References

  1. Alvim FC, Carolino SMB, Cascardo JCM, Nunes CC, Martinez CA, Otoni WC, Fontes EPB (2001) Enhanced accumulation of BiP in transgenic plants confers tolerance to water stress. Plant Physiol 126:1042–1054CrossRefPubMedGoogle Scholar
  2. Baumann K, De Paolis A, Constantino P, Gualberti G (1999) The DNA binding site of the Dof protein NtBBF1 is essential for tissue-specific and auxin-regulated expression of the rolB oncogene in plants. Plant Cell 11:323–334CrossRefPubMedGoogle Scholar
  3. Bustos M, Guiltinan M, Jordano J, Bergum D, Kalkan F, Hall T (1989) Regulation of B-glucuronidase expression in transgenic tobacco plants by an A/T-rich cis-acting sequence found upstream of a French bean B-phaseolin gene. Plant Cell 1:839–853CrossRefPubMedGoogle Scholar
  4. Buzeli RAA, Cascardo JCM, Rodrigues LAZ, Andrade MO, Almeida RS, Loureiro ME, Otoni WC, Fontes EPB (2002) Tissue-specific regulation of BiP genes: a cis-acting regulatory domain is required for BiP promoter activity in plant meristems. Plant Mol Biol 50:757–771CrossRefPubMedGoogle Scholar
  5. Castillo J, Rodrigo MI, Marques JA, Zuanigat AA, Franco L (2000) A pea nuclear protein that is induced by dehydration belongs to the vicilin superfamily. Eur J Biochem 267:2156–2165CrossRefPubMedGoogle Scholar
  6. Contim LAS, Waclawovsky AJ, Delú-Filho N, Pirovani CP, Clarindo WR, Loureiro ME, Carvalho CR, Fontes EPB (2003) The soybean sucrose binding protein gene family: genomic organization, gene copy number and tissue-specific expression of the SBP2 promoter. J Exp Bot 54:2643–2653CrossRefPubMedGoogle Scholar
  7. Delú-Filho N, Pirovani CP, Pedra JHF, Matrangolo FSV, Macedo JNA, Otoni WC, Fontes EPB (2000) A sucrose binding protein homologue from soybean affects sucrose uptake in transgenic tobacco suspension-cultured cells. Plant Physiol Biochem 38:353–361CrossRefGoogle Scholar
  8. Dunwell JM, Khuri S, Gane PJ (2000) Microbial relatives of the seed storage proteins of higher plants: conservation of structure and diversification of function during evolution of the cupin superfamily. Microbiol Mol Biol Rev 64:153–179CrossRefPubMedGoogle Scholar
  9. Dunwell JM, Purvis A, Khuri S (2004) Cupins: the most functionally diverse protein superfamily. Phytochemistry 65:7–17CrossRefPubMedGoogle Scholar
  10. Elmer A, Chao W, Grimes H (2003) Protein sorting and expression of a unique soybean cotyledon protein, GmSBP, destined for the protein storage vacuole. Plant Mol Biol 52:1089–1106CrossRefPubMedGoogle Scholar
  11. Gidoni D, Brosio P, Bond-Nutter D, Bedbrook J, Dunsmuir P (1989) Novel cis-acting elements in Petunia Cab gene promoters. Mol Gen Genet 215:337–344CrossRefPubMedGoogle Scholar
  12. Gowik U, Burscheidt J, Akyildiz M, Schule U, Koczor M, Streubel M, Westhoff P (2004) Cis-regulatory elements for mesophyll-specific gene expression in the C4 plant Flaveria trinervia, the promoter of the C4 phosphoenolpyruvate carboxylase gene. Plant Cell 16:1077–1090CrossRefPubMedGoogle Scholar
  13. Grimes HD, Overvoorde PJ (1996) Functional characterization of sucrose binding protein-mediated sucrose uptake in yeast. J Exp Bot 47:1217–1222Google Scholar
  14. Grimes HD, Overvoorde PJ, Ripp KG, Franceschi VR, Hitz WD (1992) A 62-kDa sucrose binding protein is expressed and localizes in tissues actively engaged in sucrose transport. Plant Cell 4:1561–1574CrossRefPubMedGoogle Scholar
  15. Hamilton DA, Schwarz YH, Mascarenhas JP (1998) A monocot pollen-specific promoter contains separable pollen-specific and quantitative elements. Plant Mol Biol 38:663–669CrossRefPubMedGoogle Scholar
  16. Heim U, Wang Q, Kurz T, Borisjuk L, Golombek S, Neubohn B, Adler K, Gahrtz M, Sauer N, Weber H, Wobus U (2001) Expression patterns and subcellular localization of a 52 kDa sucrose binding protein homologue of Vicia faba (VfSBPL) suggest different functions during development. Plant Mol Biol 47:461–474CrossRefGoogle Scholar
  17. Karimi M, Inze D, Depicker A (2002) GATEWAY vectors for Agrobacterium-mediated plant transformation. Trends Plant Sci 7:193–195CrossRefPubMedGoogle Scholar
  18. Kühn C, Quick WP, Schultz A, Sonnewald U, Frommer WB (1996) Companion cell-specific inhibition of the potato sucrose transporter SUT1. Plant Cell Environ 19:1115–1123CrossRefGoogle Scholar
  19. Kusnetsov V, Landsberger M, Meurer J, Oelmuller R (1999) The assembly of the CAAT-box binding complex at a photosynthesis gene promoter is regulated by light, cytokinin, and the stage of the plastids. J Biol Chem 274:36009–36014CrossRefPubMedGoogle Scholar
  20. Lam E, Chua NH (1989) ASF-2: a factor that binds to the cauliflower mosaic virus 35S promoter and a conserved GATA motif in cab promoters. Plant Cell 1:1147–1156CrossRefPubMedGoogle Scholar
  21. McCabe DE, Swain WF, Martinell BJ, Christou P (1988) Stable transformation of soybean (Glycine max) by particle bombardment. Biotechnology 6:923–926CrossRefGoogle Scholar
  22. Overvoorde PJ, Frommer WB, Spencer D (1996) A soybean sucrose binding protein independently mediates nonsaturable sucrose uptake in yeast. Plant Cell 8:271–280CrossRefPubMedGoogle Scholar
  23. Overvoorde PJ, Chao WS, Grimes HD (1997) A plasma membrane sucrose-binding protein that mediates sucrose uptake shares structural and sequence similarity with seed storage proteins but remains functionally distinct. J Biol Chem 272:15898–15904CrossRefPubMedGoogle Scholar
  24. Pedra JHF, Delú-Filho N, Pirovani CP, Contim LAS, Dewey RE, Otoni WC, Fontes EPB (2000) Antisense and sense expression of a sucrose binding protein homologue gene from soybean in transgenic tobacco affects plant growth and carbohydrate partitioning in leaves. Plant Sci 152:87–98CrossRefGoogle Scholar
  25. Pirovani CP, Macedo JNA, Contim LAS, Matrangolo FSV, Loureiro ME, Fontes EPB (2002) A sucrose binding protein homologue from soybean exhibits GTP-binding activity that functions independently of sucrose transport activity. Eur J Biochem 269:3998–4008CrossRefPubMedGoogle Scholar
  26. Polanco R, Lobos S, Vicuna R (2002) Binding of nuclear proteins to the promoter region of the lactase gene Cs-lcs1 from the basidiomycete Ceriporiopsis subvermispora. Enzyme Microb Technol 30:525–528CrossRefGoogle Scholar
  27. Rieping M, Schoffl F (1992) Synergistic effect of upstream sequences, CCAAT box elements, and HSE for enhances expression of chimeric heat shock genes in transgenic tobacco. Mol Gen Genet 231:226–232PubMedGoogle Scholar
  28. Riesmeir JW, Willmitzer L, Frommer WB (1994) Evidence for an essential role of the sucrose transporter in phloem loading and assimilate partitioning. EMBO J 13:1–7Google Scholar
  29. Ripp KG, Viitanen PV, Hitz WD, Fransceschi VR (1988) Identification of a membrane protein associated with sucrose transport into cells of developing soybean cotyledons. Plant Physiol 88:1435–1445CrossRefPubMedGoogle Scholar
  30. Rocha CS, Luz DF, Oliveira ML, Baracat-Pereira MC, Medrano FJ, Fontes EPB (2007) Expression of the sucrose binding protein from soybean: renaturation and stability of the recombinant protein. Phytochemistry 68:802–810CrossRefPubMedGoogle Scholar
  31. Ruiz-Rivero OJ, Prat S (1998) A -308 deletion of the tomato LAP promoters is able to direct flower-specific and MeJA-induced expression in transgenic plants. Plant Mol Biol 36:639–648CrossRefPubMedGoogle Scholar
  32. Stougaard J, Sandal NN, Gron A, Khule A, Marcker KA (1987) 5′ analysis of the soybean leghaemoglobin lbc(3) gene: regulatory elements required for promoter activity and organ specificity. EMBO J 6:3565–3569PubMedGoogle Scholar
  33. Stromvik MV, Sundeararaman VP, Vodkin LO (1999) A novel promoter from soybean that is active in a complex development pattern with and without its proximal 650 base pairs. Plant Mol Biol 41:217–231CrossRefPubMedGoogle Scholar
  34. Waclawovsky AJ, Freitas RL, Rocha CS, Contim LAS, Fontes EPB (2006a) Combinatorial regulation modules on GmSBP2 promoter: a distal cis-regulatory domain confines the SBP2 promoter to the vascular tissue in vegetative organs. Biochim Biophys Acta 1759:89–98PubMedGoogle Scholar
  35. Waclawovsky AJ, Loureiro ME, Freitas RL, Rocha CS, Cano MAO, Fontes EPB (2006b) Evidence for the sucrose-binding protein role in carbohydrate metabolism and transport at early development stage. Physiol Plant 128:391–404CrossRefGoogle Scholar
  36. Warmbrodt RD, Buckhout TJ, Hitz WD (1989) Localization of a protein, immunologically similar to a sucrose-binding protein from developing soybean cotyledons, on the plasma membrane of sieve-tube members of spinach leaves. Planta 180:105–115CrossRefGoogle Scholar
  37. Warmbrodt RD, Vanderwoude WJ, Hitz WD (1991) Studies on the localization of a protein, immunologically similar to a 62-kilodalton sucrose-binding protein isolated from developing soybean cotyledons, in the shoot and root of spinach. New Phytol 118:501–511CrossRefGoogle Scholar
  38. Yoshihara T, Washida H, Takaiwa F (1996) 45-bp proximal region containing AACA and GCN4 motif is sufficient to confer endosperm-specific expression of the rice storage protein glutein gene, GluA-3. FEBS Lett 383:213–218CrossRefPubMedGoogle Scholar
  39. Zhao Y, Leisy DJ, Okita TW (1994) Tissue-specific expression and temporal regulation of the rice glutein Gt3 gene are conferred by at least two spatially separated cis-regulatory elements. Plant Mol Biol 25:429–436CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Rejane L. Freitas
    • 1
  • Claudine M. Carvalho
    • 2
  • Luciano G. Fietto
    • 2
  • Marcelo E. Loureiro
    • 2
  • Andrea M. Almeida
    • 1
  • Elizabeth P. B. Fontes
    • 2
  1. 1.Departamento de Biologia VegetalUniversidade Federal de ViçosaVicosaBrazil
  2. 2.Departamento de Bioquímica e Biologia Molecular, BIOAGROUniversidade Federal de ViçosaVicosaBrazil

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