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Molecular details of the CPSF73-CPSF100 C-terminal heterodimer and interaction with Symplekin

Eukaryotic pre-mRNA is processed by a large multiprotein complex to accurately cleave the 3′ end, and to catalyse the addition of the poly(A) tail. Within this cleavage and polyadenylation specificity factor (CPSF) machinery, the CPSF73/CPSF3 endonuclease subunit directly contacts both CPSF100/CPSF2...

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Detalles Bibliográficos
Autores principales: Thore, Stéphane, Raoelijaona, Finaritra, Talenton, Vincent, Fribourg, Sébastien, Mackereth, Cameron D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10688271/
https://www.ncbi.nlm.nih.gov/pubmed/37989222
http://dx.doi.org/10.1098/rsob.230221
Descripción
Sumario:Eukaryotic pre-mRNA is processed by a large multiprotein complex to accurately cleave the 3′ end, and to catalyse the addition of the poly(A) tail. Within this cleavage and polyadenylation specificity factor (CPSF) machinery, the CPSF73/CPSF3 endonuclease subunit directly contacts both CPSF100/CPSF2 and the scaffold protein Symplekin to form a subcomplex known as the core cleavage complex or mammalian cleavage factor. Here we have taken advantage of a stable CPSF73-CPSF100 minimal heterodimer from Encephalitozoon cuniculi to determine the solution structure formed by the first and second C-terminal domain (CTD1 and CTD2) of both proteins. We find a large number of contacts between both proteins in the complex, and notably in the region between CTD1 and CTD2. A similarity is also observed between CTD2 and the TATA-box binding protein (TBP) domains. Separately, we have determined the structure of the terminal CTD3 domain of CPSF73, which also belongs to the TBP domain family and is connected by a flexible linker to the rest of CPSF73. Biochemical assays demonstrate a key role for the CTD3 of CPSF73 in binding Symplekin, and structural models of the trimeric complex from other species allow for comparative analysis and support an overall conserved architecture.