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PMID: 21993300 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

An essential role for Clp1 in assembly of polyadenylation complex CF IA and Pol II transcription termination.

Nucleic acids research ·Vol. 40 ·No. 3 ·2012-02-00 ·Pages 1226-39

Haddad R, Maurice F, Viphakone N, Voisinet-Hakil F, Fribourg S, Minvielle-Sébastia L

Abstract

Polyadenylation is a co-transcriptional process that modifies mRNA 3'-ends in eukaryotes. In yeast, CF IA and CPF constitute the core 3'-end maturation complex. CF IA comprises Rna14p, Rna15p, Pcf11p and Clp1p. CF IA interacts with the C-terminal domain of RNA Pol II largest subunit via Pcf11p which links pre-mRNA 3'-end processing to transcription termination. Here, we analysed the role of Clp1p in 3' processing. Clp1p binds ATP and interacts in CF IA with Pcf11p only. Depletion of Clp1p abolishes transcription termination. Moreover, we found that association of mutations in the ATP-binding domain and in the distant Pcf11p-binding region impair 3'-end processing. Strikingly, these mutations prevent not only Clp1p-Pcf11p interaction but also association of Pcf11p with Rna14p-Rna15p. ChIP experiments showed that Rna15p cross-linking to the 3'-end of a protein-coding gene is perturbed by these mutations whereas Pcf11p is only partially affected. Our study reveals an essential role of Clp1p in CF IA organization. We postulate that Clp1p transmits conformational changes to RNA Pol II through Pcf11p to couple transcription termination and 3'-end processing. These rearrangements likely rely on the correct orientation of ATP within Clp1p.

MeSH Terms
Alleles Mutation Polyadenylation Protein Subunits/metabolism RNA 3' End Processing RNA Polymerase II/metabolism RNA, Small Nucleolar/genetics Saccharomyces cerevisiae Proteins/genetics,metabolism,physiology Transcription, Genetic mRNA Cleavage and Polyadenylation Factors/genetics,metabolism,physiology
Chemicals
Clp1 protein, S cerevisiae PCF11 protein, S cerevisiae Protein Subunits RNA, Small Nucleolar Saccharomyces cerevisiae Proteins mRNA Cleavage and Polyadenylation Factors RNA Polymerase II
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Haddad Raphaël
Université Bordeaux Segalen, Bordeaux, F-33000, France.
Maurice Frédérique
Viphakone Nicolas
Voisinet-Hakil Florence
Fribourg Sébastien
Minvielle-Sébastia Lionel
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkr800
Published
2012-02-00
Epub
2011-00-12
Pages
1226-39
Language
English
Region
England
NLM ID
0411011
PMCID
PMC3273802
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkr800 PMC Full Text View journal details
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