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Structural differences in translation initiation between pathogenic trypanosomatids and their mammalian hosts

View ORCID ProfileAnthony Bochler, View ORCID ProfileJailson Brito Querido, View ORCID ProfileTerezie Prilepskaja, Heddy Soufari, Angelita Simonetti, Mayara Lucia Del Cistia, View ORCID ProfileLauriane Kuhn, View ORCID ProfileAline Rimoldi Ribeiro, Leoš Shivaya Valášek, View ORCID ProfileYaser Hashem
doi: https://doi.org/10.1101/806141
Anthony Bochler
1INSERM U1212 (ARNA), Institut Européen de Chimie et Biologie, Université de Bordeaux, Pessac 33607, France
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  • ORCID record for Anthony Bochler
Jailson Brito Querido
1INSERM U1212 (ARNA), Institut Européen de Chimie et Biologie, Université de Bordeaux, Pessac 33607, France
5CNRS UPR9002 (ARN), Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg F-67000, France
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Terezie Prilepskaja
2Laboratory of Regulation of Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, Prague, 142 20, the Czech Republic
3Charles University, Faculty of Science, Albertov 6, 128 00 Prague 2, Czech Republic
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Heddy Soufari
1INSERM U1212 (ARNA), Institut Européen de Chimie et Biologie, Université de Bordeaux, Pessac 33607, France
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Angelita Simonetti
5CNRS UPR9002 (ARN), Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg F-67000, France
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Mayara Lucia Del Cistia
1INSERM U1212 (ARNA), Institut Européen de Chimie et Biologie, Université de Bordeaux, Pessac 33607, France
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Lauriane Kuhn
4Université de Strasbourg, CNRS, Plateforme Protéomique Strasbourg-Esplanade FRC 1589, Strasbourg F-67000, France
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Aline Rimoldi Ribeiro
1INSERM U1212 (ARNA), Institut Européen de Chimie et Biologie, Université de Bordeaux, Pessac 33607, France
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Leoš Shivaya Valášek
2Laboratory of Regulation of Gene Expression, Institute of Microbiology of the Czech Academy of Sciences, Videnska 1083, Prague, 142 20, the Czech Republic
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Yaser Hashem
1INSERM U1212 (ARNA), Institut Européen de Chimie et Biologie, Université de Bordeaux, Pessac 33607, France
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  • For correspondence: yaser.hashem@inserm.fr
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SUMMARY

Canonical mRNA translation in eukaryotes begins with the formation of the 43S pre-initiation complex (PIC). Its assembly requires the binding of several eukaryotic initiation factors (eIF 1, 1A, 2, 3 and 5), Met-tRNAiMet and the small ribosomal subunit (40S). Compared to their mammalian hosts, trypanosomatids present significant structural differences in their 40S suggesting substantial variability in translation initiation. Here we determined the structure of the 43S PIC from Trypanosoma cruzi, the parasite causing the Chagas disease. Our structure shows numerous specific features, such as the variant eIF3 structure and its unique interactions with the large rRNA ESs 9S, 7S and 6S, and the association of a kinetoplastid-specific ~245 kDa DDX60-like helicase. It also revealed the so-far-elusive 40S-binding site of the eIF5 C-terminal domain and the structures of key terminal tails of several conserved eIFs underlying their activities within the PIC. Our results are corroborated by GST-pulldown assays in both human and T. cruzi and mass-spectrometry data.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted October 28, 2020.
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Structural differences in translation initiation between pathogenic trypanosomatids and their mammalian hosts
Anthony Bochler, Jailson Brito Querido, Terezie Prilepskaja, Heddy Soufari, Angelita Simonetti, Mayara Lucia Del Cistia, Lauriane Kuhn, Aline Rimoldi Ribeiro, Leoš Shivaya Valášek, Yaser Hashem
bioRxiv 806141; doi: https://doi.org/10.1101/806141
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Structural differences in translation initiation between pathogenic trypanosomatids and their mammalian hosts
Anthony Bochler, Jailson Brito Querido, Terezie Prilepskaja, Heddy Soufari, Angelita Simonetti, Mayara Lucia Del Cistia, Lauriane Kuhn, Aline Rimoldi Ribeiro, Leoš Shivaya Valášek, Yaser Hashem
bioRxiv 806141; doi: https://doi.org/10.1101/806141

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