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Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF

Most eukaryotic messenger RNAs (mRNAs) are processed at their 3′ end by the cleavage and polyadenylation specificity factor (CPF/CPSF). CPF mediates the endonucleolytic cleavage of the pre-mRNA and addition of a polyadenosine (poly(A)) tail, which together define the 3′ end of the mature transcript....

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Autores principales: Rodríguez-Molina, Juan B., O’Reilly, Francis J., Fagarasan, Holly, Sheekey, Eleanor, Maslen, Sarah, Skehel, J. Mark, Rappsilber, Juri, Passmore, Lori A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9380774/
https://www.ncbi.nlm.nih.gov/pubmed/35584695
http://dx.doi.org/10.1016/j.molcel.2022.04.021
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author Rodríguez-Molina, Juan B.
O’Reilly, Francis J.
Fagarasan, Holly
Sheekey, Eleanor
Maslen, Sarah
Skehel, J. Mark
Rappsilber, Juri
Passmore, Lori A.
author_facet Rodríguez-Molina, Juan B.
O’Reilly, Francis J.
Fagarasan, Holly
Sheekey, Eleanor
Maslen, Sarah
Skehel, J. Mark
Rappsilber, Juri
Passmore, Lori A.
author_sort Rodríguez-Molina, Juan B.
collection PubMed
description Most eukaryotic messenger RNAs (mRNAs) are processed at their 3′ end by the cleavage and polyadenylation specificity factor (CPF/CPSF). CPF mediates the endonucleolytic cleavage of the pre-mRNA and addition of a polyadenosine (poly(A)) tail, which together define the 3′ end of the mature transcript. The activation of CPF is highly regulated to maintain the fidelity of RNA processing. Here, using cryo-EM of yeast CPF, we show that the Mpe1 subunit directly contacts the polyadenylation signal sequence in nascent pre-mRNA. The region of Mpe1 that contacts RNA also promotes the activation of CPF endonuclease activity and controls polyadenylation. The Cft2 subunit of CPF antagonizes the RNA-stabilized configuration of Mpe1. In vivo, the depletion or mutation of Mpe1 leads to widespread defects in transcription termination by RNA polymerase II, resulting in transcription interference on neighboring genes. Together, our data suggest that Mpe1 plays a major role in accurate 3′ end processing, activating CPF, and ensuring timely transcription termination.
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spelling pubmed-93807742022-08-17 Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF Rodríguez-Molina, Juan B. O’Reilly, Francis J. Fagarasan, Holly Sheekey, Eleanor Maslen, Sarah Skehel, J. Mark Rappsilber, Juri Passmore, Lori A. Mol Cell Article Most eukaryotic messenger RNAs (mRNAs) are processed at their 3′ end by the cleavage and polyadenylation specificity factor (CPF/CPSF). CPF mediates the endonucleolytic cleavage of the pre-mRNA and addition of a polyadenosine (poly(A)) tail, which together define the 3′ end of the mature transcript. The activation of CPF is highly regulated to maintain the fidelity of RNA processing. Here, using cryo-EM of yeast CPF, we show that the Mpe1 subunit directly contacts the polyadenylation signal sequence in nascent pre-mRNA. The region of Mpe1 that contacts RNA also promotes the activation of CPF endonuclease activity and controls polyadenylation. The Cft2 subunit of CPF antagonizes the RNA-stabilized configuration of Mpe1. In vivo, the depletion or mutation of Mpe1 leads to widespread defects in transcription termination by RNA polymerase II, resulting in transcription interference on neighboring genes. Together, our data suggest that Mpe1 plays a major role in accurate 3′ end processing, activating CPF, and ensuring timely transcription termination. Cell Press 2022-07-07 /pmc/articles/PMC9380774/ /pubmed/35584695 http://dx.doi.org/10.1016/j.molcel.2022.04.021 Text en © 2022 MRC Laboratory of Molecular Biology https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rodríguez-Molina, Juan B.
O’Reilly, Francis J.
Fagarasan, Holly
Sheekey, Eleanor
Maslen, Sarah
Skehel, J. Mark
Rappsilber, Juri
Passmore, Lori A.
Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF
title Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF
title_full Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF
title_fullStr Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF
title_full_unstemmed Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF
title_short Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF
title_sort mpe1 senses the binding of pre-mrna and controls 3′ end processing by cpf
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9380774/
https://www.ncbi.nlm.nih.gov/pubmed/35584695
http://dx.doi.org/10.1016/j.molcel.2022.04.021
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