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

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.
Mesh Terms:
Blotting, Western, Calorimetry, Chromatography, Gel, Crystallization, DEAD-box RNA Helicases, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Exosome Multienzyme Ribonuclease Complex, Fluorescence Polarization, Models, Molecular, Multiprotein Complexes, Nuclear Proteins, Oligonucleotide Probes, Protein Conformation, RNA-Binding Proteins, Rosaniline Dyes, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins
EMBO J.
Date: Dec. 01, 2014
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