PMID:24558481

Citation	Barbosa, RL, Legrand, P, Wien, F, Pineau, B, Thompson, A and Guimarães, BG (2014) RRP6 from Trypanosoma brucei: crystal structure of the catalytic domain, association with EAP3 and activity towards structured and non-structured RNA substrates. PLoS ONE 9:e89138
Abstract	RRP6 is a 3'-5' exoribonuclease associated to the eukaryotic exosome, a multiprotein complex essential for various RNA processing and degradation pathways. In Trypanosoma brucei, RRP6 associates with the exosome in stoichiometric amounts and was localized in both cytoplasm and nucleus, in contrast to yeast Rrp6 which is exclusively nuclear. Here we report the biochemical and structural characterization of T. brucei RRP6 (TbRRP6) and its interaction with the so-called T. brucei Exosome Associated Protein 3 (TbEAP3), a potential orthologue of the yeast Rrp6 interacting protein, Rrp47. Recombinant TbEAP3 is a thermo stable homodimer in solution, however it forms a heterodimeric complex with TbRRP6 with 1∶1 stoichiometry. The crystallographic structure of the TbRRP6 catalytic core exposes for the first time the native catalytic site of this RNase and also reveals a disulfide bond linking two helices of the HRDC domain. RNA degradation assays show the distributive exoribonuclease activity of TbRRP6 and novel findings regarding the structural range of its RNA substrates. TbRRP6 was able to degrade single and double-stranded RNAs and also RNA substrates containing stem-loops including those with 3' stem-loop lacking single-stranded extensions. Finally, association with TbEAP3 did not significantly interfere with the TbRRP6 catalytic activity in vitro.
Links	PubMed PMC3928423 Online version:10.1371/journal.pone.0089138
Keywords	Base Sequence; Chromatography, Gel; Circular Dichroism; Cloning, Molecular; Electrophoretic Mobility Shift Assay; Mass Spectrometry; Models, Molecular; Molecular Sequence Data; Mutagenesis, Site-Directed; Oligonucleotides/genetics; Protein Conformation; Protozoan Proteins/chemistry; Protozoan Proteins/genetics; Sequence Alignment; Trypanosoma brucei brucei/chemistry
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:LKHA4		GO:0000175: 3'-5'-exoribonuclease activity	ECO:0000250:	UniProtKB:Q57X81	F		HHPRED shows an e value of 1.4E-13. Figure two from the article shows sequence similarity between the two proteins. Along with a gel electrophoresis comparing the bands between of dna between the two proteins	complete CACAO 11915
TRYB2:Q38AY4		GO:0005634: nucleus	ECO:0000250:	UniProtKB:Q57X81	C		Figure two from the article shows sequence similarity between the two proteins. Along with a gel electrophoresis comparing the bands between of dna between the two proteins	complete CACAO 12084

References

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