6eoj

PolyA polymerase module of the cleavage and polyadenylation factor (CPF) from Saccharomyces cerevisiaePolyA polymerase module of the cleavage and polyadenylation factor (CPF) from Saccharomyces cerevisiae

6eoj, resolution 3.55Å

Structural highlights

6eoj is a 3 chain structure with sequence from Baker's yeast. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
Ligands:
Gene:	CFT1, YHH1, YDR301W (Baker's yeast), YTH1, YPR107C (Baker's yeast), PFS2, YNL317W, N0348 (Baker's yeast)
Experimental data:	Check
Resources:	FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT

Function

[CFT1_YEAST] RNA-binding component of the cleavage and polyadenylation factor (CPF) complex, which plays a key role in polyadenylation-dependent pre-mRNA 3'-end formation and cooperates with cleavage factors including the CFIA complex and NAB4/CFIB. Involved in poly(A) site recognition. May be involved in coupling transcription termination and mRNA 3'-end formation.^[1] ^[2] [PFS2_YEAST] Integral and essential component of the cleavage and polyadenylation factor (CPF) complex, which plays a key role in polyadenylation-dependent pre-mRNA 3'-end formation and cooperates with cleavage factors including the CFIA complex and NAB4/CFIB. May bridge the CPF and CFIA complexes.^[3] [YTH1_YEAST] RNA-binding component of the cleavage and polyadenylation factor (CPF) complex, which plays a key role in polyadenylation-dependent pre-mRNA 3'-end formation and cooperates with cleavage factors including the CFIA complex and NAB4/CFIB.^[4]

Publication Abstract from PubMed

Newly transcribed eukaryotic pre-mRNAs are processed at their 3'-ends by the ~1 MDa multiprotein cleavage and polyadenylation factor (CPF). CPF cleaves pre-mRNAs, adds a poly(A) tail and triggers transcription termination but it is unclear how its different enzymes are coordinated and assembled. Here, we show that the nuclease, polymerase and phosphatase activities of yeast CPF are organized into three modules. Using cryo-EM, we determine a 3.5 A resolution structure of the ~200 kDa polymerase module. This reveals four beta propellers in an assembly strikingly similar to other protein complexes that bind nucleic acid. Combined with in vitro reconstitution experiments, our data show that the polymerase module brings together factors required for specific and efficient polyadenylation, to help coordinate mRNA 3'-end processing.

Architecture of eukaryotic mRNA 3'-end processing machinery.,Casanal A, Kumar A, Hill CH, Easter AD, Emsley P, Degliesposti G, Gordiyenko Y, Santhanam B, Wolf J, Wiederhold K, Dornan GL, Skehel M, Robinson CV, Passmore LA Science. 2017 Oct 26. pii: eaao6535. doi: 10.1126/science.aao6535. PMID:29074584^[5]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

References

↑ Dichtl B, Blank D, Sadowski M, Hubner W, Weiser S, Keller W. Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination. EMBO J. 2002 Aug 1;21(15):4125-35. PMID:12145212
↑ Stumpf G, Domdey H. Dependence of yeast pre-mRNA 3'-end processing on CFT1: a sequence homolog of the mammalian AAUAAA binding factor. Science. 1996 Nov 29;274(5292):1517-20. PMID:8929410
↑ Ohnacker M, Barabino SM, Preker PJ, Keller W. The WD-repeat protein pfs2p bridges two essential factors within the yeast pre-mRNA 3'-end-processing complex. EMBO J. 2000 Jan 4;19(1):37-47. PMID:10619842 doi:http://dx.doi.org/10.1093/emboj/19.1.37
↑ Tacahashi Y, Helmling S, Moore CL. Functional dissection of the zinc finger and flanking domains of the Yth1 cleavage/polyadenylation factor. Nucleic Acids Res. 2003 Mar 15;31(6):1744-52. PMID:12626716
↑ Casanal A, Kumar A, Hill CH, Easter AD, Emsley P, Degliesposti G, Gordiyenko Y, Santhanam B, Wolf J, Wiederhold K, Dornan GL, Skehel M, Robinson CV, Passmore LA. Architecture of eukaryotic mRNA 3'-end processing machinery. Science. 2017 Oct 26. pii: eaao6535. doi: 10.1126/science.aao6535. PMID:29074584 doi:http://dx.doi.org/10.1126/science.aao6535

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OCA

[1] Dichtl B, Blank D, Sadowski M, Hubner W, Weiser S, Keller W. Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination. EMBO J. 2002 Aug 1;21(15):4125-35. PMID:12145212

[2] Stumpf G, Domdey H. Dependence of yeast pre-mRNA 3'-end processing on CFT1: a sequence homolog of the mammalian AAUAAA binding factor. Science. 1996 Nov 29;274(5292):1517-20. PMID:8929410

[3] Ohnacker M, Barabino SM, Preker PJ, Keller W. The WD-repeat protein pfs2p bridges two essential factors within the yeast pre-mRNA 3'-end-processing complex. EMBO J. 2000 Jan 4;19(1):37-47. PMID:10619842 doi:http://dx.doi.org/10.1093/emboj/19.1.37

[4] Tacahashi Y, Helmling S, Moore CL. Functional dissection of the zinc finger and flanking domains of the Yth1 cleavage/polyadenylation factor. Nucleic Acids Res. 2003 Mar 15;31(6):1744-52. PMID:12626716

[5] Casanal A, Kumar A, Hill CH, Easter AD, Emsley P, Degliesposti G, Gordiyenko Y, Santhanam B, Wolf J, Wiederhold K, Dornan GL, Skehel M, Robinson CV, Passmore LA. Architecture of eukaryotic mRNA 3'-end processing machinery. Science. 2017 Oct 26. pii: eaao6535. doi: 10.1126/science.aao6535. PMID:29074584 doi:http://dx.doi.org/10.1126/science.aao6535

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