6I1D

Structure of the Ysh1-Mpe1 nuclease complex from S.cerevisiae

Summary for 6I1D

• Entry DOI: 10.2210/pdb6i1d/pdb
• Descriptor: Endoribonuclease YSH1, Protein MPE1, ZINC ION, ... (5 entities in total)
• Functional Keywords: pre-mrna, mrna, nuclease, endonuclease, cleavage, polyadenylation, polya, cpf, metallo-beta-lactamase, 3' ends, gene regulation
• Biological source: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); More
• Total number of polymer chains: 2
• Total formula weight: 71756.75

Authors

Hill, C.H.,Boreikaite, V.,Kumar, A.,Casanal, A.,Kubik, P.,Degliesposti, G.,Maslen, S.,Mariani, A.,von Loeffelholz, O.,Girbig, M.,Skehel, M.,Passmore, L.A. (deposition date: 2018-10-28, release date: 2019-02-13, Last modification date: 2024-01-24)

Primary citation

Hill, C.H.,Boreikaite, V.,Kumar, A.,Casanal, A.,Kubik, P.,Degliesposti, G.,Maslen, S.,Mariani, A.,von Loeffelholz, O.,Girbig, M.,Skehel, M.,Passmore, L.A.
Activation of the Endonuclease that Defines mRNA 3' Ends Requires Incorporation into an 8-Subunit Core Cleavage and Polyadenylation Factor Complex.
Mol.Cell, 73:1217-1231.e11, 2019

PubMed Abstract: Cleavage and polyadenylation factor (CPF/CPSF) is a multi-protein complex essential for formation of eukaryotic mRNA 3' ends. CPF cleaves pre-mRNAs at a specific site and adds a poly(A) tail. The cleavage reaction defines the 3' end of the mature mRNA, and thus the activity of the endonuclease is highly regulated. Here, we show that reconstitution of specific pre-mRNA cleavage with recombinant yeast proteins requires incorporation of the Ysh1 endonuclease into an eight-subunit "CPF" complex. Cleavage also requires the accessory cleavage factors IA and IB, which bind substrate pre-mRNAs and CPF, likely facilitating assembly of an active complex. Using X-ray crystallography, electron microscopy, and mass spectrometry, we determine the structure of Ysh1 bound to Mpe1 and the arrangement of subunits within CPF. Together, our data suggest that the active mRNA 3' end processing machinery is a dynamic assembly that is licensed to cleave only when all protein factors come together at the polyadenylation site.

PubMed: 30737185
DOI: 10.1016/j.molcel.2018.12.023

Experimental method

X-RAY DIFFRACTION (2.28 Å)