Structural basis for 5′ -end-specific recognition of guide RNA by the A. fulgidus Piwi protein Journal Article
| Authors: | Ma, J. B. ; Yuan, Y. R.; Meister, G.; Pei, Y.; Tuschl, T.; Patel, D. J. |
| Article Title: | Structural basis for 5′ -end-specific recognition of guide RNA by the A. fulgidus Piwi protein |
| Abstract: | RNA interference (RNAi) is a conserved sequence-specific gene regulatory mechanism mediated by the RNA-induced silencing complex (RISC), which is composed of a single-stranded guide RNA and an Argonaute protein. The PIWI domain, a highly conserved motif within Argonaute, has been shown to adopt an RNase H fold critical for the endonuclease cleavage activity of RISC. Here we report the crystal structure of Archaeoglobus fulgidus Piwi protein bound to double-stranded RNA, thereby identifying the binding pocket for guide-strand 5′-end recognition and providing insight into guide-strand-mediated messenger RNA target recognition. The phosphorylated 5′ end of the guide RNA is anchored within a highly conserved basic pocket, supplemented by the carboxy-terminal carboxylate and a bound divalent cation. The first nucleotide from the 5′ end of the guide RNA is unpaired and stacks over a conserved tyrosine residue, whereas successive nucleotides form a four-base-pair RNA duplex. Mutation of the corresponding amino acids that contact the 5′ phosphate in human Ago2 resulted in attenuated mRNA cleavage activity. Our structure of the Piwi-RNA complex, and that determined elsewhere, provide direct support for the 5′ region of the guide RNA serving as a nucleation site for pairing with target mRNA and for a fixed distance separating the RISC-mediated mRNA cleavage site from the anchored 5′ end of the guide RNA. |
| Keywords: | unclassified drug; gene mutation; genetics; nonhuman; molecular genetics; mutant protein; protein domain; protein motif; proteins; metabolism; genes; small interfering rna; rna interference; rna; bacterial protein; chemistry; amino acid sequence; molecular sequence data; messenger rna; rna, messenger; nucleotide sequence; substrate specificity; base sequence; binding site; crystal structure; models, molecular; protein structure, tertiary; binding sites; chemical structure; conformation; nucleic acid conformation; crystallization; structure analysis; enzyme specificity; argonaute protein; phosphate; phosphates; protein tertiary structure; molecular model; double stranded rna; archaeal proteins; rna, double-stranded; nucleation; protein rna binding; piwi protein; ribonuclease h; initiation factor; archaeal protein; peptide initiation factors; positive ions; argonaute proteins; divalent cations; target recognition; ago2 protein; eukaryotic initiation factor 2c 2, human; eukaryotic initiation factor 2c-2, human; guide rna; archaeoglobus fulgidus; rna, guide |
| Journal Title: | Nature |
| Volume: | 434 |
| Issue: | 7033 |
| ISSN: | 0028-0836 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2005-03-31 |
| Start Page: | 666 |
| End Page: | 670 |
| Language: | English |
| DOI: | 10.1038/nature03514 |
| PUBMED: | 15800629 |
| PROVIDER: | scopus |
| PMCID: | PMC4694588 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 255" - "Export Date: 24 October 2012" - "CODEN: NATUA" - "Source: Scopus" |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work