The gene family encoding the Arabidopsis thaliana translation elongation factor EF-1α: Molecular cloning, characterization and expression

Abstract

The gene family encoding the Arabidopsis thaliana translation elongation factor (EF-1 alpha) was analysed. This family contains four genes (A1-A4) organized in a similar manner in different varieties of Arabidopsis. Based upon both their physical separation and a comparison of their sequences, it is suggested that the A4 gene and the A1, A2, and A3 genes constitute two distinct subfamilies within the genome. By introducing chimaeric gene constructs into Arabidopsis cells, we showed that the A1 gene promoter mediates a transient expression about twofold higher than that obtained using the CaMV 35 S promoter. This expression depends on a 348 bp DNA fragment extending from -982 to -634 bp upstream of the initiation codon. This element contains a characteristic telomeric sequence (AACCCTAA) which is also found in the promoters of the A2 and A4 genes as well as in the promoters of the Drosophila EF-1 alpha F1 gene and of several highly expressed plant genes.

A calmodulin-sensitive interaction between microtubules and a higher plant homolog of elongation factor-1 alpha.

“…In tomato (Pokalsky et al, 1989), tobacco (Ursin et al, 1991;Marty et al, 1993), and possibly Arabidopsis (Curie et al, 1993), elevated levels of EF-lu gene expression were indicated in developing tissues. Auxin (Ursin et al, 1991), low temperature (Dunn et al, 1993), and light (Aguilar et al, 1991) appear to modulate expression of some EF-lu genes, and multigene families are present in severa1 plants (Axelos et al, 1989;Pokalsky et al, 1989;Aguilar et al, 1991;Dunn et al, 1993). A gene family has been found in carrot cells as well (Kawahara et al, 1992), and Figure 46 presents data consistent with this finding.…”

Expression of the Arabidopsis HMG2 gene, encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase, is restricted to meristematic and floral tissues.

“…Protoplasts were prepared from an Arabidopsis thaliana (Columbia ecotype) cell suspension culture (lhe T87) according to Axelos et al (1989). Cells were grown on MS medium supplemented with 1 pM naphthaleneacetic acid (Axelos et al, 1989) and collected at the midexponential growth phase.…”

“…Cells were grown on MS medium supplemented with 1 pM naphthaleneacetic acid (Axelos et al, 1989) and collected at the midexponential growth phase. Transfections were performed according to the Ca(NO&polyethylene glycol technique as described by Prols et al (1988) using 40 pg of cesium chloride-purified plasmid DNA and 107 protoplasts.…”

Rhizobium meliloti elicits transient expression of the early nodulin gene ENOD12 in the differentiating root epidermis of transgenic alfalfa.

“…Because of the presente of a second Ncol site within the MtENOD12 promoter (Figure 2), it was necessary to carry out a partial digestion of pUC19-prMtl2 after linearization with EcoRl to recover the full-length 23-kb EcoRI-Ncol fragment. This promoter fragment was then cloned between the EcoRl and Ncol polylinker sites of pCCOGUS (Axelos et al, 1989) so that subsequent digestion with EcoRl and Pstl would yield a DNA fragment containing the MtENOD12 promoter fused to the gusA coding sequence with a 3' flanking polyadenylation signal (cauliflower mosaic virus 35s). After adding an Sstl site to the 3'terminus, the resultant fragment was cloned between the unique EcoRl and Sstl sites of the binary vector pLPl00 to give pLP100-prMt12.…”