Translation initiation factors eIF4E and eIFiso4E are required for polysome formation and regulate plant growth in tobacco.
Eukaryotic initiation factor eIF4E plays a pivotal role in translation initiation. As a component of the ternary eIF4F complex, eIF4E interacts with the mRNA cap structure to facilitate recruitment of the 40S ribosomal subunit onto mRNA. Plants contain two distinct cap-binding proteins, eIF4E and eIFiso4E, that assemble into different eIF4F complexes. To study the functional roles of eIF4E and eIFiso4E in tobacco, we isolated two corresponding cDNAs, NteIF4E1 and NteIFiso4E1, and used these to deplete cap-binding protein levels in planta by antisense downregulation. Antibodies raised against recombinant NteIF4E1 detected three distinct cap-binding proteins in tobacco leaf extracts; NteIF4E and two isoforms of NteIFiso4E. The three cap-binding proteins were immuno-detected in all tissues analysed and were coordinately regulated, with peak expression in anthers and pollen. Transgenic tobacco plants showing significant depletion of either NteIF4E or the two NteIFiso4E isoforms displayed normal vegetative development and were fully fertile. Interestingly, NteIFiso4E depletion resulted in a compensatory increase in NteIF4E levels, whereas the down-regulation of NteIF4E did not trigger a reciprocal increase in NteIFiso4E levels. The antisense depletion of both NteIF4E and NteIFiso4E resulted in plants with a semi-dwarf phenotype and an overall reduction in polyribosome loading, demonstrating that both eIF4E and eIFiso4E support translation initiation in planta, which suggests their potential role in the regulation of plant growth.
Department of Biology, University of Leicester, LE1 7RH Leicester, United Kingdom.
Jonathan P
JP
Combe
Marie E
ME
Petracek
Gerben
G
van Eldik
Frank
F
Meulewaeter
David
D
Twell
D000595:Amino Acid Sequence; D015153:Blotting, Western; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D015536:Down-Regulation; D039561:Eukaryotic Initiation Factor-4E / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D010802:Phylogeny; D030821:Plants, Genetically Modified; D011132:Polyribosomes / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D016373:DNA, Antisense; D018076:DNA, Complementary; D039561:Eukaryotic Initiation Factor-4E; D020033:Protein Isoforms