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EMBO J. 2014 Dec 1;33(23):2829-46. doi: 10.15252/embj.201488757. Epub 2014 Oct 15.

The exosome-binding factors Rrp6 and Rrp47 form a composite surface for recruiting the Mtr4 helicase.

Author information

1
Structural Cell Biology Department, Max Planck Institute of Biochemistry, Martinsried, Germany.
2
Molecular Biology and Biotechnology Department, The University of Sheffield, Sheffield, UK.
3
Molecular Biology and Biotechnology Department, The University of Sheffield, Sheffield, UK p.j.mitchell@sheffield.ac.uk conti@biochem.mpg.de.
4
Structural Cell Biology Department, Max Planck Institute of Biochemistry, Martinsried, Germany p.j.mitchell@sheffield.ac.uk conti@biochem.mpg.de.

Abstract

The exosome is a conserved multi-subunit ribonuclease complex that functions in 3' end processing, turnover and surveillance of nuclear and cytoplasmic RNAs. In the yeast nucleus, the 10-subunit core complex of the exosome (Exo-10) physically and functionally interacts with the Rrp6 exoribonuclease and its associated cofactor Rrp47, the helicase Mtr4 and Mpp6. Here, we show that binding of Mtr4 to Exo-10 in vitro is dependent upon both Rrp6 and Rrp47, whereas Mpp6 binds directly and independently of other cofactors. Crystallographic analyses reveal that the N-terminal domains of Rrp6 and Rrp47 form a highly intertwined structural unit. Rrp6 and Rrp47 synergize to create a composite and conserved surface groove that binds the N-terminus of Mtr4. Mutation of conserved residues within Rrp6 and Mtr4 at the structural interface disrupts their interaction and inhibits growth of strains expressing a C-terminal GFP fusion of Mtr4. These studies provide detailed structural insight into the interaction between the Rrp6-Rrp47 complex and Mtr4, revealing an important link between Mtr4 and the core exosome.

KEYWORDS:

RNA degradation; X‐ray crystallography; nuclear exosome; yeast genetics

PMID:
25319414
PMCID:
PMC4282559
DOI:
10.15252/embj.201488757
[Indexed for MEDLINE]
Free PMC Article

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