Plant Molecular Biology

, Volume 29, Issue 5, pp 995–1004 | Cite as

Definition of constitutive gene expression in plants: the translation initiation factor 4A gene as a model

  • Therese Mandel
  • Andrew J. Fleming
  • Rolf Krähenbühl
  • Cris Kuhlemeier
Research Article


The NeIF-4A10 gene belongs to a family of at least ten genes, all of which encode closely related isoforms of translation initiation factor 4A. The promoter region of NeIF-4A10 was sequenced, and four mRNA 5′ ends were determined. Deletions containing 2750, 689 and 188 bp of untranscribed upstream DNA were fused to the GUS reporter gene and introduced into transgenic tobacco. The three constructs mediated GUS expression in all cells of the leaf, stem and shoot apical meristem. Control experiments using in situ hybridization and tissue printing indicated that the observed GUS expression matches the expression patterns of NeIF-4A mRNA and protein. This detailed analysis at the level of mRNA, protein and reporter gene expression shows that NeIF-4A10 is an ideal constitutively expressed control gene. We argue that inclusion of such a control gene in experiments dealing with specifically expressed genes is in many cases essential for the correct interpretation of observed expression patterns.

Key words

cell size constitutive gene expression histochemistry in situ hybridization shoot apical meristem transgenic plants 


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  1. 1.
    Aguilar F, Montandon P-E, Stutz E: Two genes encoding the soybean translation elongation factor eEF-1α are transcribed in seedling leaves. Plant Mol Biol 17: 351–360 (1991).Google Scholar
  2. 2.
    Angenent GC, Franken J, Busscher M, Colombo L, van Tunen AJ: Petal and stamen formation in petunia is regulated by the homeotic gene fbp1. Plant J 4: 101–112 (1993).Google Scholar
  3. 3.
    Avni D, Shama S, Loreni F, Meyuhas O: Vertebrate mRNAs with a 5′ terminal pyrimidine tract are candidates for translational repression in quiescent cells: characterization of the translational cis-regulatory element. Mol Cell Biol 14: 3822–3833 (1994).Google Scholar
  4. 4.
    Benfey PN, Ren L, Chua N-H: Combinatorial and synergistic properties of CaMV 35S enhancer domains. EMBO J 9: 1677–1696 (1990).Google Scholar
  5. 5.
    Brander KA, Kuhlemeier C: A pollen-specific DEAD-box protein related to translation initiation factor eIF-4A. Plant Mol Biol 27: 637–649 (1995).Google Scholar
  6. 6.
    Brander KA, Mandel T, Owttrim GW, Kuhlemeier C: Highly conserved genes coding for eukaryotic translation initiation factor eIF-4A of tobacco have specific alterations in functional motifs. Biochim Biophys Acta 1261: 442–444 (1995).Google Scholar
  7. 7.
    Bucher M, Kuhlemeier C: Long term anoxia tolerance: multi-level regulation of gene expression in the amphibious plant Acorus calamus L. Plant Physiol 103: 441–448 (1993).Google Scholar
  8. 8.
    Curie C, Liboz T, Bardet C, Gander E, Medal C, Axelos M, Lescure B: Cis and trans-acting elements involved in the activation of Arabidopsis thaliana A1 gene encoding the translation elongation factor EF-1. Nucl Acids Res 19: 1305–1310 (1991).Google Scholar
  9. 9.
    Curie C, Axelos M, Bardet C, Atanassova R, Chaubet N, Lescure B: Modular organization and developmental activity of an Arabidopsis thaliana EF-1 gene promoter. Mol Gen. Genet 238: 428–436 (1993).Google Scholar
  10. 10.
    Fleming AJ, Mandel T, Hofmann S, Sterk P, deVries SC, Kuhlemeier C: Expression pattern of a tobacco lipid transfer protein gene within the shoot apex. Plant J 2: 855–862 (1992).Google Scholar
  11. 11.
    Fleming AJ, Mandel T, Roth I, Kuhlemeier C: The patterns of gene expression in the tomato shoot apical meristem. Plant Cell 5: 297–309 (1993).Google Scholar
  12. 12.
    Jefferies HBJ, Reinhard C, Kozma SC, Thomas G: Rapamycin selectively represses translation of the ‘polypyrimidine tract’ mRNA family. Proc Natl Acad Sci USA 91: 4441–4445 (1994).Google Scholar
  13. 13.
    Jefferson RA, Kavanagh T, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).Google Scholar
  14. 14.
    Jorgensen RA: Cosuppression, flower color patterns, and metastable gene expression states. Science 268: 686–691 (1995).Google Scholar
  15. 15.
    Kohler S, Coraggio I, Becker D, Salamini F: Pattern of meristem-specific cDNA clones of barley (Hordeum vulgare L). Planta 186: 227–235 (1992).Google Scholar
  16. 16.
    Kuhlemeier C, Fluhr R, Green PJ, Chua NH: Sequences in the pea rbcS-3A gene have homology to constitutive mammalian enhancers but function as negative regulatory elements. Genes Devel 1: 247–255 (1987).Google Scholar
  17. 17.
    Kuhlemeier C, Green PJ, Chua N-H: Regulation of gene expression in plants. Annu Rev Plant Physiol 38: 221–257 (1987).Google Scholar
  18. 18.
    Lu J, Lee W, Jiang C, Keller EB: Start site selection by Sp1 in the TATA-less human Ha-ras promoter. J Biol Chem 269: 5391–5402 (1994).Google Scholar
  19. 19.
    Medford JI, Elmer JS, Klee HJ: Molecular cloning and characterization of genes expressed in shoot apical meristems. Plant Cell 3: 359–370 (1991).Google Scholar
  20. 20.
    Merrick WC: Mechanisms and regulation of eukaryotic protein synthesis. Microbiol Rev 56: 291–315 (1992).Google Scholar
  21. 21.
    Owttrim GW, Hofmann S, Kuhlemeier C: Divergent genes for translation initiation factor eIF-4A are coordinately expressed in tobacco. Nucl Acids Res 19: 5491–5496 (1991).Google Scholar
  22. 22.
    Owttrim GW, Mandel T, Trachsel H, Thomas AAM, Kuhlemeier C: Characterization of the tobacco eIF-4A gene family. Plant Mol Biol 26: 1747–1757 (1994).Google Scholar
  23. 23.
    Pokalski AR, Hiatt WR, Ridge N, Rasmussen R, Houck CM, Shewmaker CK: Structure and expression of elongation factor 1α in tomato. Nucl Acids Res 17: 4661–4673 (1989).Google Scholar
  24. 24.
    Ryser U, Keller B: Ultrastructural localization of a bean glycine-rich protein in unlignified primary walls of protoxylem cells. Plant Cell 4: 773–783 (1992).Google Scholar
  25. 25.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  26. 26.
    Sijmons PC, Dekker BMM, Schrammeijer B, Verwoerd TC, Van denElzen PJM, Hoekema A: Production of correctly processed human serum albumin in transgenic plants. Bio/technology 8: 217–221 (1990).Google Scholar
  27. 27.
    Smale ST, Baltimore D: The ‘initiator’ as a transcription control element. Cell 57: 103–113 (1989).Google Scholar
  28. 28.
    Taylor CB, Bariola PA, DelCardayre SB, Raines RT, Green PJ: RNS2: a senescence-associated RNase of Arabidopsis that diverged from the S-RNases before speciation. Proc Natl Acad Sci USA 90: 5118–5122 (1993).Google Scholar
  29. 29.
    Terada N, Patel HR, Takase K, Kohno K, Nairn A, Gelfand EW: Rapamycin selectively inhibits translation of mRNAs encoding elongation factors and ribosomal proteins. Proc Natl Acad Sci USA 91: 11477–11481 (1994).Google Scholar
  30. 30.
    Thoma S, Hecht U, Kippers A, Botella J, deVries S, Somerville C: Tissue-specific expression of a gene encoding a cell wall-localized lipid transfer protein. Plant Physiol 105: 35–45 (1994).Google Scholar
  31. 31.
    Uknes S, Dincher S, Friedrich L, Negrotto D, Williams S, Thompson-Taylor H, Potter S, Ward E, Ryals J: Regulation of pathogenesis-related protein la expression in tobacco. Plant Cell 5: 159–169 (1993).Google Scholar
  32. 32.
    VanLijsebettens M, Vanderhaeghen R, DeBlock M, Bauw G, Villaroel R, VanMontagu M: An S18 ribosomal protein gene copy at the Arabidopsis PFL locus affects plant development by its specific expression in meristems. EMBO J 13: 3378–3388 (1994).Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Therese Mandel
    • 1
  • Andrew J. Fleming
    • 1
  • Rolf Krähenbühl
    • 1
  • Cris Kuhlemeier
    • 1
  1. 1.Institute of Plant PhysiologyUniversity of BerneBerneSwitzerland

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