Functional insights from a surface antigen mRNA-bound proteome

Abstract
Data availability
Article and author information
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Abstract

Trypanosoma brucei is the causative agent of human sleeping sickness. The parasites' Variant Surface Glycoprotein (VSG) enables them to evade adaptive immunity via antigenic variation. VSG comprises 10% of total cell protein and the high stability of VSG mRNA is essential for trypanosome survival. To determine how VSG mRNA stability is maintained, we used mRNA affinity purification to identify all its associated proteins. CFB2, an unconventional RNA-binding protein with an F-box domain, was specifically enriched with VSG mRNA. We demonstrate that CFB2 is essential for VSG mRNA stability, describe cis acting elements within the VSG 3'-untranslated region that regulate the interaction, identify trans-acting factors that are present in the VSG messenger ribonucleoprotein particle and mechanistically explain how CFB2 stabilizes the mRNA of this key pathogenicity factor. Beyond T. brucei, the mRNP purification approach has the potential to supply detailed biological insight into metabolism of relatively abundant mRNAs in any eukaryote.

Data availability

The RNASeq raw data is available at Array Express with the accession number E-MTAB-9700. The proteomics data are available via ProteomeXchange with identifier PXD021772.

The following data sets were generated

1. Clayton C
2. Erben E
(2020) Effect of CFB2 depletion in Trypanosoma brucei
ArrayExpress, E-MTAB-9700.

https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-9700/
1. Clayton C
2. Erben E
(2020) The trypanosome Variant Surface Glycoprotein mRNA is stabilized by an essential unconventional RNA-binding protein
ProteomeXchange, PXD021772.

http://proteomecentral.proteomexchange.org/cgi/GetDataset?ID=PXD021772

Article and author information

Author details

Larissa Melo do Nascimento

CC lab, ZMBH, Heidelberg, Germany

Competing interests
No competing interests declared.
Franziska Egler

CC lab, ZMBH, Heidelberg, Germany

Competing interests
No competing interests declared.
Katharina Arnold

CC lab, ZMBH, Heidelberg, Germany

Competing interests
No competing interests declared.
Nina Papavasiliou

Deutsche Krebsforschungszentrum (DKFZ), Heidelberg, Germany

Competing interests
No competing interests declared.
Christine Clayton

Zentrum fuer Molekulare Biologie der Universitaet Heidelberg, DKFZ-ZMBH Alliance, Heidelberg, Germany

Competing interests
Christine Clayton, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0002-6384-0731
Esteban Erben

Molecular Parasitology, IIBIO - UNSAM, San Martín, Argentina

For correspondence
eerben@iib.unsam.edu.ar

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-5179-7863

Funding

H2020 European Research Council (649019)

Nina Papavasiliou

Core Funding University of Heidelberg (ND)

Christine Clayton

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.