Arabidopsis ARGONAUTE1 is an RNA Slicer that selectively recruits microRNAs and short interfering RNAs. Export

@article{Baumberger2005, abstract = {ARGONAUTE (AGO) RNA-binding proteins are involved in RNA silencing. They bind to short interfering RNAs (siRNAs) and microRNAs (miRNAs) through a conserved PAZ domain, and, in animals, they assemble into a multisubunit RNA-induced silencing complex (RISC). The mammalian AGO2, termed Slicer, directs siRNA- and miRNA-mediated cleavage of a target RNA. In Arabidopsis, there are 10 members of the AGO family, and the AGO1 protein is potentially the Slicer component in different RNA-silencing pathways. Here, we show that AGO1 selectively recruits certain classes of short silencing-related RNA. AGO1 is physically associated with miRNAs, transacting siRNAs, and transgene-derived siRNAs but excludes virus-derived siRNAs and 24-nt siRNAs involved in chromatin silencing. We also show that AGO1 has Slicer activity. It mediates the in vitro cleavage of a mir165 target RNA in a manner that depends on the sequence identity of amino acid residues in the PIWI domain that are predicted by homology with animal Slicer-competent AGO proteins to constitute the RNase catalytic center. However, unlike animals, we find no evidence that AGO1 Slicer is in a high molecular weight RNA-induced silencing complex. The Slicer activity fractionates as a complex of approximately 150 kDa that likely constitutes the AGO1 protein and associated RNA without any other proteins. Based on sequence similarity, we predict that other Arabidopsis AGOs might have a similar catalytic activity but recruit different subsets of siRNAs or miRNAs.}, address = {Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, United Kingdom.}, author = {Baumberger, N. and Baulcombe, D. C.}, citeulike-article-id = {399108}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0505461102}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/16081530}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=16081530}, day = {16}, doi = {10.1073/pnas.0505461102}, issn = {0027-8424}, journal = {Proc Natl Acad Sci U S A}, keywords = {silencing, tesi}, month = {August}, number = {33}, pages = {11928--11933}, posted-at = {2007-12-19 11:44:13}, priority = {4}, title = {Arabidopsis ARGONAUTE1 is an RNA Slicer that selectively recruits microRNAs and short interfering RNAs.}, url = {http://dx.doi.org/10.1073/pnas.0505461102}, volume = {102}, year = {2005} }

TY - JOUR ID - Baumberger2005 L3 - citeulike-article-id:399108 N2 - ARGONAUTE (AGO) RNA-binding proteins are involved in RNA silencing. They bind to short interfering RNAs (siRNAs) and microRNAs (miRNAs) through a conserved PAZ domain, and, in animals, they assemble into a multisubunit RNA-induced silencing complex (RISC). The mammalian AGO2, termed Slicer, directs siRNA- and miRNA-mediated cleavage of a target RNA. In Arabidopsis, there are 10 members of the AGO family, and the AGO1 protein is potentially the Slicer component in different RNA-silencing pathways. Here, we show that AGO1 selectively recruits certain classes of short silencing-related RNA. AGO1 is physically associated with miRNAs, transacting siRNAs, and transgene-derived siRNAs but excludes virus-derived siRNAs and 24-nt siRNAs involved in chromatin silencing. We also show that AGO1 has Slicer activity. It mediates the in vitro cleavage of a mir165 target RNA in a manner that depends on the sequence identity of amino acid residues in the PIWI domain that are predicted by homology with animal Slicer-competent AGO proteins to constitute the RNase catalytic center. However, unlike animals, we find no evidence that AGO1 Slicer is in a high molecular weight RNA-induced silencing complex. The Slicer activity fractionates as a complex of approximately 150 kDa that likely constitutes the AGO1 protein and associated RNA without any other proteins. Based on sequence similarity, we predict that other Arabidopsis AGOs might have a similar catalytic activity but recruit different subsets of siRNAs or miRNAs. IS - 33 JF - Proc Natl Acad Sci U S A SN - 0027-8424 AD - Sainsbury Laboratory, Norwich Research Park, Norwich NR4 7UH, United Kingdom. EP - 11933 TI - Arabidopsis ARGONAUTE1 is an RNA Slicer that selectively recruits microRNAs and short interfering RNAs. VL - 102 SP - 11928 KW - silencing KW - tesi AU - Baumberger, N AU - Baulcombe, DC PY - 2005/08/16/ UR - http://dx.doi.org/10.1073/pnas.0505461102 ER -