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.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Haddad R,Maurice F,Viphakone N,Voisinet-Hakil F,Fribourg S,Minvielle-Sébastia Ldoi
10.1093/nar/gkr800subject
Has Abstractpub_date
2012-02-01 00:00:00pages
1226-39issue
3eissn
0305-1048issn
1362-4962pii
gkr800journal_volume
40pub_type
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