Translational initiation in Leishmania tarentolae and Phytomonas serpens (Kinetoplastida) is strongly influenced by pre-ATG triplet and its 5' sequence context

Molecular and Biochemical Parasitology
Julius LukesKirill Alexandrov

Abstract

To investigate the influence of sequence context of translation initiation codon on translation efficiency in Kinetoplastida, we constructed a library of expression plasmids randomized in the three nucleotides prefacing ATG of a reporter gene encoding enhanced green fluorescent protein (EGFP). All 64 possible combinations of pre-ATG triplets were individually stably integrated into the rDNA locus of Leishmania tarentolae and the resulting cell lines were assessed for EGFP expression. The expression levels were quantified directly by measuring the fluorescence of EGFP protein in living cells and confirmed by Western blotting. We observed a strong influence of the pre-ATG triplet on the level of protein expression over a 20-fold range. To understand the degree of evolutionary conservation of the observed effect, we transformed Phytomonas serpens, a trypanosomatid parasite of plants, with a subset of the constructs. The pattern of translational efficiency mediated by individual pre-ATG triplets in this species was similar to that observed in L. tarentolae. However, the pattern of translational efficiency of two other proteins (red fluorescent protein and tetracycline repressor) containing selected pre-ATG triplets did not correlat...Continue Reading

Citations

Aug 7, 2007·Parasitology Research·Cristina Folgueira, Jose M Requena
Sep 26, 2009·Molecular Biotechnology·Giancarlo Basile, Manuela Peticca
Apr 24, 2009·Journal of Biomolecular Structure & Dynamics·Silvie Foldynová-TrantirkováLukás Trantírek
Oct 4, 2012·Parasites & Vectors·Alsu ChajbullinovaPetr Volf
Sep 4, 2007·Molecular and Biochemical Parasitology·Christine Clayton, Michal Shapira
Jan 8, 2008·Current Opinion in Microbiology·Simon Haile, Barbara Papadopoulou
Nov 19, 2017·Nature Communications·Moran Shalev-BenamiGeorgios Skiniotis
Mar 23, 2019·Parasite : Journal De La Société Française De Parasitologie·Petra ČermákováAnton Horváth
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Oct 23, 2020·Nucleic Acids Research·Donna MatzovMoran Shalev-Benami
Jan 8, 2021·Parasitology·Petra ČermákováAnton Horváth

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