7NH2

Crystal structure of Toxoplasma CPSF4-YTH domain bound to m6A

Summary for 7NH2

• Entry DOI: 10.2210/pdb7nh2/pdb
• Related: 7NG2
• Descriptor: Zinc finger (CCCH type) motif-containing protein, TETRAETHYLENE GLYCOL, ~{N},9-dimethylpurin-6-amine, ... (4 entities in total)
• Functional Keywords: yth, cleavage and polyadenylation, rna, m6a, rna binding protein
• Biological source: Toxoplasma gondii (strain ATCC 50611 / Me49)
• Total number of polymer chains: 1
• Total formula weight: 18805.40

Authors

Swale, C.,Bowler, M.W. (deposition date: 2021-02-09, release date: 2021-07-21, Last modification date: 2024-01-31)

Primary citation

Farhat, D.C.,Bowler, M.W.,Communie, G.,Pontier, D.,Belmudes, L.,Mas, C.,Corrao, C.,Coute, Y.,Bougdour, A.,Lagrange, T.,Hakimi, M.A.,Swale, C.
A plant-like mechanism coupling m6A reading to polyadenylation safeguards transcriptome integrity and developmental gene partitioning in Toxoplasma .
Elife, 10:-, 2021

PubMed 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 of an m6A-dependent 3'end polyadenylation serving as a transcriptional barrier at these . We identify the core polyadenylation complex within 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 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 is associated with an increase in aberrantly elongated chimeric mRNAs emanating from impaired transcriptional termination, a phenotype previously noticed in the plant model . 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 and . 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 and , as a guardian against aberrant transcriptional read-throughs.

PubMed: 34263725
DOI: 10.7554/eLife.68312

Experimental method

X-RAY DIFFRACTION (1.35 Å)