Multimeric assembly and biochemical characterization of the Trax-translin endonuclease complex Journal Article
| Authors: | Tian, Y.; Simanshu, D. K.; Ascano, M.; Diaz-Avalos, R.; Park, A. Y.; Juranek, S. A.; Rice, W. J.; Yin, Q.; Robinson, C. V.; Tuschl, T.; Patel, D. J. |
| Article Title: | Multimeric assembly and biochemical characterization of the Trax-translin endonuclease complex |
| Abstract: | Trax-translin heteromers, also known as C3PO, have been proposed to activate the RNA-induced silencing complex (RISC) by facilitating endonucleolytic cleavage of the siRNA passenger strand. We report on the crystal structure of hexameric Drosophila C3PO formed by truncated translin and Trax, along with electron microscopic and mass spectrometric studies on octameric C3PO formed by full-length translin and Trax. Our studies establish that Trax adopts the translin fold, possesses catalytic centers essential for C3PO's endoRNase activity and interacts extensively with translin to form an octameric assembly. The catalytic pockets of Trax subunits are located within the interior chamber of the octameric scaffold. Truncated C3PO, like full-length C3PO, shows endoRNase activity that leaves 3-2-hydroxyl-cleaved ends. We have measured the catalytic activity of C3PO and shown it to cleave almost stoichiometric amounts of substrate per second. © 2011 Nature America, Inc. All rights reserved. |
| Keywords: | unclassified drug; mass spectrometry; electron microscopy; animals; microscopy, electron; protein assembly; enzyme activity; rna; kinetics; protein multimerization; carrier proteins; crystal structure; models, molecular; crystallography, x-ray; drosophila melanogaster; molecular interaction; catalysis; catalytic domain; protein structure, quaternary; drosophila proteins; stoichiometry; endoribonucleases; ribonuclease; multiprotein complex; component 3 promoter of rna induced silencing complex |
| Journal Title: | Nature Structural and Molecular Biology |
| Volume: | 18 |
| Issue: | 6 |
| ISSN: | 1545-9993 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2011-06-01 |
| Start Page: | 658 |
| End Page: | 664 |
| Language: | English |
| DOI: | 10.1038/nsmb.2069 |
| PROVIDER: | scopus |
| PMCID: | PMC3109869 |
| PUBMED: | 21552261 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 17 August 2011" - "CODEN: NSMBC" - "Source: Scopus" |
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