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

Lukes, Julius, Paris, Zdeněk, Regmi, Sandesh, Breitling, Reinhard, Mureev, Sergey, Kushnir, Susanna, Pyatkov, Konstantin, Jirků, Milan and Alexandrov, Kirill A. (2006) 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, 148 2: 125-132. doi:10.1016/j.molbiopara.2006.03.008


Author Lukes, Julius
Paris, Zdeněk
Regmi, Sandesh
Breitling, Reinhard
Mureev, Sergey
Kushnir, Susanna
Pyatkov, Konstantin
Jirků, Milan
Alexandrov, Kirill A.
Title Translational initiation in Leishmania tarentolae and Phytomonas serpens (Kinetoplastida) is strongly influenced by pre-ATG triplet and its 5' sequence context
Journal name Molecular and Biochemical Parasitology   Check publisher's open access policy
ISSN 0166-6851
Publication date 2006-08-01
Year available 2006
Sub-type Article (original research)
DOI 10.1016/j.molbiopara.2006.03.008
Open Access Status
Volume 148
Issue 2
Start page 125
End page 132
Total pages 8
Place of publication Amsterdam
Publisher Elsevier/North Holland
Language eng
Subject 060199 Biochemistry and Cell Biology not elsewhere classified
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 correlate with either EGFP or each other. Thus, we conclude that a conserved mechanism of translation initiation site selection exists in kinetoplastids that is strongly influenced not only by the pre-ATG sequences but also by the coding region of the gene.
Keyword Translation
Initiation
Pre-ATG
Kinetoplastida
Leishmania
Q-Index Code C1
Institutional Status Non-UQ
Additional Notes PMID: 16644031

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
Institute for Molecular Bioscience - Publications
 
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