Skip to main content
SpringerLink
Log in

A pollen-specific DEAD-box protein related to translation initiation factor eIF-4A from tobacco

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

A pollen-specific sequence, NeIF-4A8, has been isolated from a cDNA library from mature pollen of Nicotiana tabacum cv. Samsun. NeIF-4A8 is a full-length cDNA whose deduced amino acid sequence exhibits high homology to the eucaryotic translation initiation factor eIF-4A from mouse, Drosophila and tobacco. eIF-4A is an RNA helicase which belongs to the supergene family of DEAD-box proteins. Northern blot analysis with a gene-specific probe showed strict anther-specific expression of NeIF-4A8 starting at microspore mitosis. With antibodies raised against tobacco eIF-4A the presence of abundant eIF-4A-related proteins in developing anthers and pollen grains was demonstrated. The genomic analysis shows that the coding region is split by three introns whereas a large, fourth intron is situated in the 5′-untranslated region. A promoter construct with 2137 bp of upstream sequence fused to the GUS reporter gene was used to confirm that the expression is confined to the haploid cells within the anther. NeIF-4A8 is a prime candidate for mediating translational control in the developing male gametophyte.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Aguilar F, Montandon P-E, Stutz E: Two genes encoding the soybean translation elongation factor eEF-1 are transcribed in seedling leaves. Plant Mol Biol 17: 351–360 (1991).

    Google Scholar 

  2. Audet RG, Goodchild J, Richter JD: Eukaryotic initiation factor 4A stimulates translation in microinjected Xenopus oocytes. Devel Biol 121: 58–68 (1987).

    Google Scholar 

  3. Bashe D, Mascarenhas JP: Changes in potassium ion concentrations during pollen dehydration and germination in relation to protein synthesis. Plant Sci Lett 35: 55–60 (1984).

    Article  Google Scholar 

  4. Brander KA, Mandel T, Owttrim GW, Kuhlemeier C: Highly conserved genes coding for eucaryotic translation initiation factor eIF-4A of tobacco have specific alterations in functional motifs. Biochim Biophys Acta, in press (1995).

  5. 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).

    Article  PubMed  Google Scholar 

  6. Capkova-Balatkova V, Hrabetova E, Tupy J: Effects of cycloheximide on pollen of Nicotiana tabacum in culture. Biochem Physiol Pflanzen 175: 412–420 (1980).

    Google Scholar 

  7. Chapman GP: The tapetum. In: Giles KL, Prakash J (eds) International Review of Cytology, pp. 111–125. Academic Press, London (1987).

    Google Scholar 

  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).

    PubMed  Google Scholar 

  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).

    Article  PubMed  Google Scholar 

  10. DeBlock M, Debrouwer D: In-situ enzyme histochemistry on plastic-embedded plant material. The development of an artefact-free β-glucuronidase assay. Plant J 2: 261–266 (1992).

    Google Scholar 

  11. Draper J, Scott R, Armitage P, Walden R: Plant Genetic Transformation and Gene Expression. Blackwell Scientific Publications, Oxford (1988).

    Google Scholar 

  12. Hu C, Chee PP, Chesney RH, Zhou JH, Miller PD, O'Brien WT: Intrinsic GUS-like activities in seed plants. Plant Cell Rep 9: 1–5 (1990).

    Article  Google Scholar 

  13. Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).

    PubMed  Google Scholar 

  14. Koltunow AM, Truettner J, Cox KH, Wallroth M, Goldberg RB: Different temporal and spatial gene expression patterns occur during anther development. Plant Cell 2: 1201–1224 (1990).

    Article  PubMed  Google Scholar 

  15. Lasko PF, Ashburner M: The product of the Drosophila gene vasa is very similar to eukaryotic initiation factor-4A. Nature 335: 611–617 (1988).

    Article  PubMed  Google Scholar 

  16. Leroy P, Alzari P, Sassoon D, Wolgemuth D, Fellous M: The protein encoded by a murine male germ line-specific transcript is a putative ATP-dependent RNA helicase. Cell 57: 549–559 (1989).

    Article  PubMed  Google Scholar 

  17. Martin T, Wöhner RV, Hummel S, Willmitzer L, Frommer WB: The GUS reporter system as a tool to study plant gene expression. In: Gallagher SR (ed) GUS Protocols, pp. 23–43. Academic Press, San Diego (1992).

    Google Scholar 

  18. Mascarenhas JP: The biochemistry of angiosperm pollen development. Bot Rev 41: 260–314 (1975).

    Google Scholar 

  19. Mascarenhas JP, Mermelstein J: Messenger RNAs: their utilization and degradation during pollen germination and tube growth. Acta Soc Bot Pol 50: 13–20 (1981).

    Google Scholar 

  20. Mascarenhas JP: The male gametophyte of flowering plants. Plant Cell 1: 657–664 (1989).

    Article  PubMed  Google Scholar 

  21. Mascarenhas JP: Gene activity during pollen development. Ann Rev Physiol Plant Mol Biol 41: 317–338 (1990).

    Article  Google Scholar 

  22. Mascarenhas JP, Hamilton DA: Artefacts in the localization of GUS activity in anthers of petunia transformed with a CaMV 35S-GUS construct. Plant J 2: 405–408 (1992).

    Google Scholar 

  23. Mascarenhas JP: Anther- and pollen-expressed genes. In: Verma DPS, Goldberg RB (eds) Temporal and Spatial Regulation of Plant Genes, pp. 97–115. Springer-Verlag, Wien (1988).

    Google Scholar 

  24. Mascarenhas NT, Bashe D, Eisenberg A, Willing RP, Xiao CM, Mascarenhas JP: Messenger RNAs in corn pollen and protein synthesis during germination and pollen tube growth. Theor Appl Genet 68: 323–326 (1984).

    Article  Google Scholar 

  25. McCormick S: Male gametophyte development. Plant Cell 5: 1265–1275 (1993).

    Article  PubMed  Google Scholar 

  26. Merrick WC: Mechanisms and regulation of eukaryotic protein synthesis. Microbiol Rev 56: 291–315 (1992).

    PubMed  Google Scholar 

  27. Nielsen PJ, McMaster GK, Trachsel H: Cloning of eucaryotic protein synthesis initiation factor genes: isolation and characterization of cDNA clones encoding factor eIF-4A. Nucl Acids Res 13: 6867–6880 (1985).

    PubMed  Google Scholar 

  28. 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).

    PubMed  Google Scholar 

  29. Owttrim GW, Mandel T, Trachsel H, Thomas AAM, Kuhlemeier C: Characterization of the tobacco eIF-4A gene family. Plant Mol Biol, in press (1994).

  30. Plegt L, Bino RJ: β-Glucuronidase activity during development of the male gametophyte from transgenic and non-transgenic plants. Mol Gen Genet 216: 321–327 (1989).

    Article  Google Scholar 

  31. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  32. Schmid SR, Linder P: D-E-A-D protein family of putative RNA helicases. Mol Microbiol 6: 283–292 (1992).

    PubMed  Google Scholar 

  33. Schrauwen JAM, deGroot PFM, vanHerpen MMA, van derLee T, Reynen WH, Weterings KAP, Wullems GJ: Stage-related expression of mRNAs during pollen development in lily and tobacco. Planta 182: 298–304 (1990).

    Article  Google Scholar 

  34. Sonenberg N, Guertin D, Lee KAW: Capped mRNAs with reduced secondary structure can function in extracts from poliovirus-infected cells. Mol Cell Biol 2: 1633–1638 (1982).

    PubMed  Google Scholar 

  35. Sood PP: Histoenzymological compartementation of β-glucuronidase in the germinating pollen grains of Protulaca grandiflora. Biol Plant 22: 124–127 (1980).

    Google Scholar 

  36. Stinson JR, Eisenberg AJ, Willing RP, Pe ME, Hanson DD, Mascarenhas JP: Genes expressed in the male gametophyte of flowering plants and their isolation. Plant Physiol 83: 442–447 (1987).

    Google Scholar 

  37. Twell D, Klein TM, Fromm ME, McCormick S: Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiol 91: 1270–1274 (1989).

    Google Scholar 

  38. Twell D, Yamaguchi J, Wing RA, Ushiba J, McCormick S: Promoter analysis of genes that are corrdinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Devel 5: 496–507 (1991).

    PubMed  Google Scholar 

  39. Twell D: The diversity and regulation of gene expression in the pathway of male gametophyte development. In: Scott R, Stead T (eds) Molecular and Cellular Basis of Plant Reproduction. Company of Biologists, Cambridge, in press (1994).

    Google Scholar 

  40. Uknes S, Dincher S, Friedrich L, Negrotto D, Williams S, Thompson-Taylor H, Potter S, Ward E, Ryals J: Regulation of pathogenesis-related protein-1a gene expression in tobacco. Plant Cell 5: 159–169 (1993).

    Article  PubMed  Google Scholar 

  41. Webster C, Gaut RL, Browning KS, Ravel JM, Roberts JKM: Hypoxia enhances phosphorylation of eukaryotic initiation factor 4A in maize root tips. J Biol Chem 266: 23341–23346 (1991).

    PubMed  Google Scholar 

  42. Zarsky V, Capkova V, Hrabetova E, Tupy J: Protein changes during pollen development in Nicotiana tabacum L. Biol Plant 27: 438–444 (1985).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Plant Physiology, University of Berne, Altenbergrain 21, CH-3013, Berne, Switzerland

    Karl A. Brander & Cris Kuhlemeier

Authors
  1. Karl A. Brander
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cris Kuhlemeier
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Brander, K.A., Kuhlemeier, C. A pollen-specific DEAD-box protein related to translation initiation factor eIF-4A from tobacco. Plant Mol Biol 27, 637–649 (1995). https://doi.org/10.1007/BF00020219

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020219

Key words

Search

Navigation