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A plant-like mechanism coupling m6A reading to polyadenylation safeguards transcriptome integrity and developmental gene partitioning in Toxoplasma

DOI: 10.7554/eLife.68312 DOI Help

Authors: Dayana C. Farhat (Université Grenoble Alpes) , Matthew W. Bowler (European Molecular Biology Laboratory) , Guillaume Communie (Institut Laue-Langevin) , Dominique Pontier (UMR5096, CNRS) , Lucid Belmudes (BIG-BGE, Université Grenoble Alpes) , Caroline Mas (Integrated Structural Biology Grenoble (ISBG), Université Grenoble Alpes) , Charlotte Corrao (Université Grenoble Alpes) , Yohann Couté (Université Grenoble Alpes) , Alexandre Bougdour (Université Grenoble Alpes) , Thierry Lagrange (UMR5096, CNRS) , Mohamed-Ali Hakimi (Université Grenoble Alpes) , Christopher Swale (Université Grenoble Alpes)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Elife , VOL 10

State: Published (Approved)
Published: July 2021
Diamond Proposal Number(s): 21969

Open Access Open Access

Abstract: Correct 3'end processing of mRNAs is one of the regulatory cornerstones of gene expression. In a parasite that must adapt to the regulatory requirements of its multi-host life style, there is a need to adopt additional means to partition the distinct transcriptional signatures of the closely and tandemly-arranged stage specific genes. In this study, we report our findings in T. gondii of an m6A-dependent 3'end polyadenylation serving as a transcriptional barrier at these loci. We identify the core polyadenylation complex within T. gondii and establish CPSF4 as a reader for m6A-modified mRNAs, via a YTH domain within its C-terminus, a feature which is shared with plants. We bring evidence of the specificity of this interaction both biochemically, and by determining the crystal structure at high resolution of the T. gondii CPSF4-YTH in complex with an m6A modified RNA. We show that the loss of m6A, both at the level of its deposition or its recognition was associated with an increase in aberrantly elongated chimeric mRNAs emanating from impaired transcriptional termination, a phenotype previously noticed in the plant model Arabidopsis thaliana. Nanopore direct RNA sequencing shows the occurrence of transcriptional read-through breaching into downstream repressed stage-specific genes, in the absence of either CPSF4 or the m6A RNA methylase components in both T. gondii and A. thaliana. Taken together, our results shed light on an essential regulatory mechanism coupling the pathways of m6A metabolism directly to the cleavage and polyadenylation processes, one that interestingly seem to serve, in both T. gondii and A. thaliana, as a guardian against aberrant transcriptional read-throughs.

Diamond Keywords: Toxoplasmosis

Subject Areas: Biology and Bio-materials

Instruments: I04-Macromolecular Crystallography

Added On: 21/07/2021 09:27

Discipline Tags:

Infectious Diseases Disease in the Developing World Health & Wellbeing Genetics Structural biology Life Sciences & Biotech Parasitology

Technical Tags:

Diffraction Macromolecular Crystallography (MX)