Architecture of eukaryotic mRNA 3′-end processing machinery

DOI: 10.1126/science.aao6535

Authors: Ana Casanal (MRC Laboratory of Molecular Biology) , Ananthanarayanan Kumar (MRC Laboratory of Molecular Biology) , Chris H. Hill (MRC Laboratory of Molecular Biology) , Ashley D. Easter (MRC Laboratory of Molecular Biology) , Paul Emsley (MRC Laboratory of Molecular Biology) , Gianluca Degliesposti (MRC Laboratory of Molecular Biology) , Yuliya Gordiyenko (MRC Laboratory of Molecular Biology; University of Oxford) , Balaji Santhanam (MRC Laboratory of Molecular Biology) , Jana Wolf (MRC Laboratory of Molecular Biology) , Katrin Wiederhold (MRC Laboratory of Molecular Biology) , Gillian L. Dornan (MRC Laboratory of Molecular Biology) , Mark Skehel (MRC Laboratory of Molecular Biology) , Carol V. Robinson (University of Oxford) , Lori A. Passmore (MRC Laboratory of Molecular Biology)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Science , VOL 546

State: Published (Approved)
Published: October 2017
Diamond Proposal Number(s): 15622

Abstract: Newly transcribed eukaryotic pre-mRNAs are processed at their 3′ ends by the ~1-megadalton multiprotein cleavage and polyadenylation factor (CPF). CPF cleaves pre-mRNAs, adds a polyadenylate tail, and triggers transcription termination, but it is unclear how its various enzymes are coordinated and assembled. Here, we show that the nuclease, polymerase, and phosphatase activities of yeast CPF are organized into three modules. Using electron cryomicroscopy, we determined a 3.5-angstrom-resolution structure of the ~200-kilodalton polymerase module. This revealed four beta-propellers, in an assembly markedly similar to those of 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 messenger RNA 3′-end processing

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: Krios I-Titan Krios I at Diamond

Other Facilities: MRC LMB

Added On: 15/11/2017 10:58

Documents:
science.aao6535.full.pdf

Discipline Tags:

Structural biology Life Sciences & Biotech

Technical Tags:

Microscopy Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM)