{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T03:29:06Z","timestamp":1767842946630,"version":"3.49.0"},"reference-count":30,"publisher":"Proceedings of the National Academy of Sciences","issue":"23","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2008,6,10]]},"abstract":"<jats:p>Small RNAs triggering RNA silencing are loaded onto Argonautes and then sequence-specifically guide them to target transcripts. Epitope-tagged human Argonautes (hAgo1, hAgo2, hAgo3, and hAgo4) are associated with siRNAs and miRNAs, but only epitope-tagged hAgo2 has been shown to have Slicer activity. Contrarily, how endogenous hAgos behave with respect to small RNA association and target RNA destruction has remained unclear. Here, we produced monoclonal antibodies for individual hAgos. High-throughput pyrosequencing revealed that immunopurified endogenous hAgo2 and hAgo3 associated mostly with miRNAs. Endogenous hAgo3 did not show Slicer function but localized in P-bodies, suggesting that hAgo3 endogenously expressed is, like hAgo2, involved in the miRNA pathway but antagonizes the RNAi activity of hAgo2. Sequence variations of miRNAs were found at both 5\u2032 and 3\u2032 ends, suggesting that multiple mature miRNAs containing different \u201cseed\u201d sequences can arise from one miRNA precursor. The hAgo antibodies we raised are valuable tools for ascertaining the functional behavior of endogenous Argonautes and miRNAs in RNA silencing.<\/jats:p>","DOI":"10.1073\/pnas.0800334105","type":"journal-article","created":{"date-parts":[[2008,6,5]],"date-time":"2008-06-05T01:20:59Z","timestamp":1212628859000},"page":"7964-7969","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":210,"title":["Characterization of endogenous human Argonautes and their miRNA partners in RNA silencing"],"prefix":"10.1073","volume":"105","author":[{"given":"Asuka","family":"Azuma-Mukai","sequence":"first","affiliation":[{"name":"Institute for Genome Research, University of Tokushima, Tokushima 770-8503, Japan;"}]},{"given":"Hideo","family":"Oguri","sequence":"additional","affiliation":[{"name":"First Institute of New Drug Discovery, Otsuka Pharmaceutical Co., Ltd., Tokushima 771-0192, Japan;"}]},{"given":"Toutai","family":"Mituyama","sequence":"additional","affiliation":[{"name":"Computational Biology Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Tokyo 135-00664, Japan;"}]},{"given":"Zhi Rong","family":"Qian","sequence":"additional","affiliation":[{"name":"Institute for Genome Research, University of Tokushima, Tokushima 770-8503, Japan;"}]},{"given":"Kiyoshi","family":"Asai","sequence":"additional","affiliation":[{"name":"Computational Biology Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Tokyo 135-00664, Japan;"},{"name":"Graduate School of Frontier Science, University of Tokyo, Chiba 277-8561, Japan;"}]},{"given":"Haruhiko","family":"Siomi","sequence":"additional","affiliation":[{"name":"Institute for Genome Research, University of Tokushima, Tokushima 770-8503, Japan;"},{"name":"Keio University School of Medicine, Tokyo 160-8582, Japan"}]},{"given":"Mikiko C.","family":"Siomi","sequence":"additional","affiliation":[{"name":"Institute for Genome Research, University of Tokushima, Tokushima 770-8503, Japan;"},{"name":"Japan Science and Technology Agency, Core Research for Evolutional Science and Technology, Saitama 332-0012, Japan; and"},{"name":"Keio University School of Medicine, Tokyo 160-8582, Japan"}]}],"member":"341","published-online":{"date-parts":[[2008,6,10]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.02.016"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1284105"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2006.09.009"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.cellbio.23.090506.123406"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.05.001"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.03.028"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1146484"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1073827"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.974702"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1102513"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2004.07.007"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.4161\/rna.4.2.4640"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1265"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.10.7960-7971.1998"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1274"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1115079"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2005.10.031"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2007.04.040"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.7179508"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(03)01018-3"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.862301"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1454806"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1140494"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1261\/rna.744307"},{"key":"e_1_3_3_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2005.08.044"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.embor.7400637"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb1308"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1565607"},{"key":"e_1_3_3_29_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.129.3.551"},{"key":"e_1_3_3_30_2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1370605"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.0800334105","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T20:59:33Z","timestamp":1649797173000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.0800334105"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,6,10]]},"references-count":30,"journal-issue":{"issue":"23","published-print":{"date-parts":[[2008,6,10]]}},"alternative-id":["10.1073\/pnas.0800334105"],"URL":"https:\/\/doi.org\/10.1073\/pnas.0800334105","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,6,10]]},"assertion":[{"value":"2008-01-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-06-10","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}