Abstract
Eukaryotic elongation factor 1A1 (eEF1A1) is central to translational activity. It is involved in complexes that form signal transduction with protein kinase C, as well as being a signal transducer and activator of transcription 3. eEF1A1 and eEF1A2 are isoforms of the alpha subunit of elongating factor 1 complex. It has been reported that eEF1A1 is expressed in most human tissues but the brain, skeletal muscle and heart. eEF1A1 has been linked to both apoptosis and anti-apoptotic activities. In this study, eEF1A1 was co-expressed with Bax, a proapoptotic protein via heterologous expression of recombinant DNA in yeast cells. Assays were carried out to monitor the fate and state of yeast cells when eEF1A1 was co-expressed with Bax. The yeast strain (bearing an integrated copy of the Bax gene) was transformed with an episomal 2-micron plasmid that encodes HA-tagged eEF1A1 gene. The resultant strain would allow co-expression of Bax and eEF1A1 in yeast cells, Bax being under the control of the GAL1 promoter, while the PGK1 promoter drives eEF1A1 expression. Bcl 2A1, a known anti-apoptotic protein, was also co-expressed with Bax in yeast cells as a positive control, to study the anti-apoptotic characteristic of eEF-1A1. The part eEF1A1 plays in apoptosis has been contentious, amidst the pro and anti-apoptotic properties of eEF1A1, it was shown clearly, in this study that eEF1A1 portrays only anti-apoptotic property in the presence of pro-apoptotic protein, Bax.
Highlights
Eukaryotic elongation factor 1A proteins have multiple functions, which include aminoacyl-tRNAs recruitment during protein synthesis to ribosomes, actin building, phosphoinositol-4 kinase activation, and microtubule splitting [1]
The resultant strain Bax(LEU2)::Eukaryotic elongation factor 1A1 (eEF1A1) would allow co-expression of Bax and eEF1A1 in yeast cells, Bax being under the control of the GAL1 promoter, while the PGK1 promoter drives eEF1A1 expression
This was similar for Bcl 2A1 (Fig. 1e, f), Fig. 1a, b show that eEF1A1 supports the growth of yeast cells carrying the toxic Bax gene when the expression of the pro-apoptotic protein Bax was fully induced in the presence of galactose
Summary
Eukaryotic elongation factor 1A proteins have multiple functions, which include aminoacyl-tRNAs recruitment during protein synthesis to ribosomes, actin building, phosphoinositol-4 kinase activation, and microtubule splitting [1]. It was reported that human eEF1A1 might naturally form oligomers, and its overexpression has been linked to the transcription of muscle pro-apoptotic and proteolysis genes in hypercatabolic trauma patients [1]. EEF1A1 protein is widely expressed in different tissues while eEF1A2 is found mainly in the brain, skeletal muscle and heart [3]. According to Bosutti et al eEF1A1 is expressed in all human tissues excepting the brain, skeletal muscle and heart. Its levels are gradually reduced at the primary phase of growth and development. It seems that eEF1A2 takes over the function of the synthesis of proteins from eEF1A1 in the adult muscle [4]. The two eEF1A isoforms act in opposite directions, during
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