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Cited by 6RNA Research and SplicingRNA and protein synthesis mechanismsProtein Structure and Dynamics

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

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Thore, Stéphane, Raoelijaona, Finaritra, Talenton, Vincent, Fribourg, Sébastien, Mackereth, Cameron D.·2023·Open Biology
DOI 10.1098/rsob.230221PMC PMC10688271

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Abstract

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.

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Metadata

Authors
Thore, S., Raoelijaona, F., Talenton, V., Fribourg, S., & Mackereth, C. D.
Affiliations
Centre National de la Recherche Scientifique (FR)
Corresponding author
  • Fribourg, Sébastien
  • Mackereth, Cameron D.
ORCID
Venue
Open Biology
Publisher
The Royal Society
Published
2023-11
Volume
13
Issue
11
Pages
230221-230221
ISSN
2046-2441 (ISSN-L)
DOI
10.1098/rsob.230221
PMC
PMC10688271
OpenAlex
W4388880407
Cited by
6
References
2
Citations by year
2026: 4 · 2024: 1 · 2023: 1
FWCI
0.53
Type
journal article
Language
en
Open access
Licence
Locations
  • Open Biology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • HAL (submittedVersion)
  • HAL (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Institut National de la Santé et de la Recherche Médicale
  • Centre National de la Recherche Scientifique
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
CPSF73, symplekin, cleavage and polyadenylation specificity factor, C-terminal domain, solution structure, Encephalitozoon cuniculi, protein-protein interaction, solution NMR
MeSH terms
Encephalitozoon cuniculi, mRNA Cleavage and Polyadenylation Factors, Cleavage And Polyadenylation Specificity Factor

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