Mutations in the Chromodomain-like Insertion of Translation Elongation Factor 3 Compromise Protein Synthesis Through Reduced ATPase Activity
Journal of Biological Chemistry
Translation elongation is mediated by ribosomes and multiple soluble factors, many of which are conserved across bacteria and eukaryotes. During elongation, eukaryotic elongation factor 1A (eEF1A; EF-Tu in bacteria) delivers aminoacylated-tRNA to the A-site of the ribosome, whereas eEF2 (EF-G in bacteria) translocates the ribosome along the mRNA. Fungal translation elongation is striking in its absolute requirement for a third factor, the ATPase eEF3. eEF3 binds close to the E-site of the ribosome and has been proposed to facilitate the removal of deacylated tRNA from the E-site. eEF3 has two ATP binding cassette (ABC) domains, the second of which carries a unique chromodomain-like insertion hypothesized to play a significant role in its binding to the ribosome. This model was tested in the current study using a mutational analysis of the Sac7d region of the chromodomain-like insertion. Specific mutations in this domain result in reduced growth rate as well as slower translation elongation. In vitro analysis demonstrates that these mutations do not affect the ability of eEF3 to interact with the ribosome. Kinetic analysis revealed a larger turnover number for ribosomes in comparison to eEF3, indicating that the partial reactions involving the ribosome are significantly faster than that of eEF3. Mutations in the chromodomain-like insertion severely compromise the ribosome stimulated ATPase of eEF3, strongly suggesting that it exerts an allosteric effect on the hydrolytic activity of eEF3. The chromodomain-like insertion is, therefore, vital to eEF3 function and may be targeted for developing novel antifungal drugs.
Sasikumar, Arjun N. and Kinzy, Terri Goss, "Mutations in the Chromodomain-like Insertion of Translation Elongation Factor 3 Compromise Protein Synthesis Through Reduced ATPase Activity" (2014). Faculty Publications – Biological Sciences. 70.
This article was originally published as Sasikumar, A.N. and Kinzy, T.G. (2014) Mutations in the Chromodomain-like Insertion of Translation Elongation Factor 3 Compromise Protein Synthesis Through Reduced ATPase Activity. J Biol Chem. 289:4853-60. PMC3931047