Phosphorylation of human Argonaute proteins affects small RNA binding
Argonaute (Ago) proteins are highly conserved between species and constitute a direct-binding platform for small RNAs including short-interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi interacting RNAs (piRNAs). Small RNAs function as guides whereas Ago proteins are the actual mediators of gene...
Gespeichert in:
| Veröffentlicht in: | Nucleic acids research 2011-03, Vol.39 (6), p.2330-2343 |
|---|---|
| Hauptverfasser: | , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 2343 |
|---|---|
| container_issue | 6 |
| container_start_page | 2330 |
| container_title | Nucleic acids research |
| container_volume | 39 |
| creator | Rüdel, Sabine Wang, Yanli Lenobel, René Körner, Roman Hsiao, He-Hsuan Urlaub, Henning Patel, Dinshaw Meister, Gunter |
| description | Argonaute (Ago) proteins are highly conserved between species and constitute a direct-binding platform for small RNAs including short-interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi interacting RNAs (piRNAs). Small RNAs function as guides whereas Ago proteins are the actual mediators of gene silencing. Although the major steps in RNA-guided gene silencing have been elucidated, not much is known about Ago-protein regulation. Here we report a comprehensive analysis of Ago2 phosphorylation in human cells. We find that the highly conserved tyrosine Y529, located in the small RNA 5'-end-binding pocket of Ago proteins can be phosphorylated. By substituting Y529 with a negatively charged glutamate (E) mimicking a phosphorylated tyrosine, we show that small RNA binding is strongly reduced. Our data suggest that a negatively charged phospho-tyrosine generates a repulsive force that prevents efficient binding of the negatively charged 5' phosphate of the small RNA. |
| doi_str_mv | 10.1093/nar/gkq1032 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3064767</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>899157436</sourcerecordid><originalsourceid>FETCH-LOGICAL-c478t-23bece11ac8f31095f1897f776c2b961efbd37f15534755521e8f40d0567f8fe3</originalsourceid><addsrcrecordid>eNqFkUtLw0AUhQdRtD5W7iU7FxI7N_PMRijFFxQV0fUwSWbaaDLTziRC_70Ra9GVq7s4H4dz-RA6BXwJOCdjp8N4_r4CTLIdNALCs5TmPNtFI0wwSwFTeYAOY3zDGCgwuo8OMsACKJYjdP208HG58GHd6K72LvE2WfStdskkzL3TfWeSZfCdqV1MtLWm7GISW900yfPDJClqV9Vufoz2rG6iOdncI_R6c_0yvUtnj7f308ksLamQXZqRwpQGQJfSkmE7syBzYYXgZVbkHIwtKiIsMEaoYIxlYKSluMKMCyutIUfo6rt32RetqUrjuqAbtQx1q8NaeV2rv4mrF2ruPxTBnAouhoLzTUHwq97ETrV1LE3TaGd8H5XMc2CCEv4_yRnPicxgIC--yTL4GIOx2z2A1ZchNRhSG0MDffb7hS37o4R8Aihqje4</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>865693821</pqid></control><display><type>article</type><title>Phosphorylation of human Argonaute proteins affects small RNA binding</title><source>PubMed Central Free</source><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Oxford Journals Open Access Collection</source><source>Free Full-Text Journals in Chemistry</source><creator>Rüdel, Sabine ; Wang, Yanli ; Lenobel, René ; Körner, Roman ; Hsiao, He-Hsuan ; Urlaub, Henning ; Patel, Dinshaw ; Meister, Gunter</creator><creatorcontrib>Rüdel, Sabine ; Wang, Yanli ; Lenobel, René ; Körner, Roman ; Hsiao, He-Hsuan ; Urlaub, Henning ; Patel, Dinshaw ; Meister, Gunter</creatorcontrib><description>Argonaute (Ago) proteins are highly conserved between species and constitute a direct-binding platform for small RNAs including short-interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi interacting RNAs (piRNAs). Small RNAs function as guides whereas Ago proteins are the actual mediators of gene silencing. Although the major steps in RNA-guided gene silencing have been elucidated, not much is known about Ago-protein regulation. Here we report a comprehensive analysis of Ago2 phosphorylation in human cells. We find that the highly conserved tyrosine Y529, located in the small RNA 5'-end-binding pocket of Ago proteins can be phosphorylated. By substituting Y529 with a negatively charged glutamate (E) mimicking a phosphorylated tyrosine, we show that small RNA binding is strongly reduced. Our data suggest that a negatively charged phospho-tyrosine generates a repulsive force that prevents efficient binding of the negatively charged 5' phosphate of the small RNA.</description><identifier>ISSN: 0305-1048</identifier><identifier>EISSN: 1362-4962</identifier><identifier>DOI: 10.1093/nar/gkq1032</identifier><identifier>PMID: 21071408</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Argonaute 2 protein ; Argonaute Proteins ; Data processing ; Eukaryotic Initiation Factor-2 - chemistry ; Eukaryotic Initiation Factor-2 - genetics ; Eukaryotic Initiation Factor-2 - metabolism ; Eukaryotic Initiation Factors - chemistry ; Eukaryotic Initiation Factors - genetics ; Eukaryotic Initiation Factors - metabolism ; Gene silencing ; Glutamic acid ; HEK293 Cells ; Humans ; Mimicry ; miRNA ; Mutation ; Phosphate ; Phosphorylation ; Phosphotyrosine - chemistry ; Protein Binding ; Protein Structure, Tertiary ; Ribonuclease III - metabolism ; RNA ; RNA, Small Interfering - metabolism ; RNA, Small Untranslated - metabolism ; RNA-Binding Proteins - metabolism ; siRNA ; Tyrosine ; Tyrosine - metabolism</subject><ispartof>Nucleic acids research, 2011-03, Vol.39 (6), p.2330-2343</ispartof><rights>The Author(s) 2010. Published by Oxford University Press. 2010</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c478t-23bece11ac8f31095f1897f776c2b961efbd37f15534755521e8f40d0567f8fe3</citedby><cites>FETCH-LOGICAL-c478t-23bece11ac8f31095f1897f776c2b961efbd37f15534755521e8f40d0567f8fe3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3064767/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3064767/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21071408$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rüdel, Sabine</creatorcontrib><creatorcontrib>Wang, Yanli</creatorcontrib><creatorcontrib>Lenobel, René</creatorcontrib><creatorcontrib>Körner, Roman</creatorcontrib><creatorcontrib>Hsiao, He-Hsuan</creatorcontrib><creatorcontrib>Urlaub, Henning</creatorcontrib><creatorcontrib>Patel, Dinshaw</creatorcontrib><creatorcontrib>Meister, Gunter</creatorcontrib><title>Phosphorylation of human Argonaute proteins affects small RNA binding</title><title>Nucleic acids research</title><addtitle>Nucleic Acids Res</addtitle><description>Argonaute (Ago) proteins are highly conserved between species and constitute a direct-binding platform for small RNAs including short-interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi interacting RNAs (piRNAs). Small RNAs function as guides whereas Ago proteins are the actual mediators of gene silencing. Although the major steps in RNA-guided gene silencing have been elucidated, not much is known about Ago-protein regulation. Here we report a comprehensive analysis of Ago2 phosphorylation in human cells. We find that the highly conserved tyrosine Y529, located in the small RNA 5'-end-binding pocket of Ago proteins can be phosphorylated. By substituting Y529 with a negatively charged glutamate (E) mimicking a phosphorylated tyrosine, we show that small RNA binding is strongly reduced. Our data suggest that a negatively charged phospho-tyrosine generates a repulsive force that prevents efficient binding of the negatively charged 5' phosphate of the small RNA.</description><subject>Argonaute 2 protein</subject><subject>Argonaute Proteins</subject><subject>Data processing</subject><subject>Eukaryotic Initiation Factor-2 - chemistry</subject><subject>Eukaryotic Initiation Factor-2 - genetics</subject><subject>Eukaryotic Initiation Factor-2 - metabolism</subject><subject>Eukaryotic Initiation Factors - chemistry</subject><subject>Eukaryotic Initiation Factors - genetics</subject><subject>Eukaryotic Initiation Factors - metabolism</subject><subject>Gene silencing</subject><subject>Glutamic acid</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Mimicry</subject><subject>miRNA</subject><subject>Mutation</subject><subject>Phosphate</subject><subject>Phosphorylation</subject><subject>Phosphotyrosine - chemistry</subject><subject>Protein Binding</subject><subject>Protein Structure, Tertiary</subject><subject>Ribonuclease III - metabolism</subject><subject>RNA</subject><subject>RNA, Small Interfering - metabolism</subject><subject>RNA, Small Untranslated - metabolism</subject><subject>RNA-Binding Proteins - metabolism</subject><subject>siRNA</subject><subject>Tyrosine</subject><subject>Tyrosine - metabolism</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtLw0AUhQdRtD5W7iU7FxI7N_PMRijFFxQV0fUwSWbaaDLTziRC_70Ra9GVq7s4H4dz-RA6BXwJOCdjp8N4_r4CTLIdNALCs5TmPNtFI0wwSwFTeYAOY3zDGCgwuo8OMsACKJYjdP208HG58GHd6K72LvE2WfStdskkzL3TfWeSZfCdqV1MtLWm7GISW900yfPDJClqV9Vufoz2rG6iOdncI_R6c_0yvUtnj7f308ksLamQXZqRwpQGQJfSkmE7syBzYYXgZVbkHIwtKiIsMEaoYIxlYKSluMKMCyutIUfo6rt32RetqUrjuqAbtQx1q8NaeV2rv4mrF2ruPxTBnAouhoLzTUHwq97ETrV1LE3TaGd8H5XMc2CCEv4_yRnPicxgIC--yTL4GIOx2z2A1ZchNRhSG0MDffb7hS37o4R8Aihqje4</recordid><startdate>20110301</startdate><enddate>20110301</enddate><creator>Rüdel, Sabine</creator><creator>Wang, Yanli</creator><creator>Lenobel, René</creator><creator>Körner, Roman</creator><creator>Hsiao, He-Hsuan</creator><creator>Urlaub, Henning</creator><creator>Patel, Dinshaw</creator><creator>Meister, Gunter</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QO</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>20110301</creationdate><title>Phosphorylation of human Argonaute proteins affects small RNA binding</title><author>Rüdel, Sabine ; Wang, Yanli ; Lenobel, René ; Körner, Roman ; Hsiao, He-Hsuan ; Urlaub, Henning ; Patel, Dinshaw ; Meister, Gunter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-23bece11ac8f31095f1897f776c2b961efbd37f15534755521e8f40d0567f8fe3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Argonaute 2 protein</topic><topic>Argonaute Proteins</topic><topic>Data processing</topic><topic>Eukaryotic Initiation Factor-2 - chemistry</topic><topic>Eukaryotic Initiation Factor-2 - genetics</topic><topic>Eukaryotic Initiation Factor-2 - metabolism</topic><topic>Eukaryotic Initiation Factors - chemistry</topic><topic>Eukaryotic Initiation Factors - genetics</topic><topic>Eukaryotic Initiation Factors - metabolism</topic><topic>Gene silencing</topic><topic>Glutamic acid</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Mimicry</topic><topic>miRNA</topic><topic>Mutation</topic><topic>Phosphate</topic><topic>Phosphorylation</topic><topic>Phosphotyrosine - chemistry</topic><topic>Protein Binding</topic><topic>Protein Structure, Tertiary</topic><topic>Ribonuclease III - metabolism</topic><topic>RNA</topic><topic>RNA, Small Interfering - metabolism</topic><topic>RNA, Small Untranslated - metabolism</topic><topic>RNA-Binding Proteins - metabolism</topic><topic>siRNA</topic><topic>Tyrosine</topic><topic>Tyrosine - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rüdel, Sabine</creatorcontrib><creatorcontrib>Wang, Yanli</creatorcontrib><creatorcontrib>Lenobel, René</creatorcontrib><creatorcontrib>Körner, Roman</creatorcontrib><creatorcontrib>Hsiao, He-Hsuan</creatorcontrib><creatorcontrib>Urlaub, Henning</creatorcontrib><creatorcontrib>Patel, Dinshaw</creatorcontrib><creatorcontrib>Meister, Gunter</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rüdel, Sabine</au><au>Wang, Yanli</au><au>Lenobel, René</au><au>Körner, Roman</au><au>Hsiao, He-Hsuan</au><au>Urlaub, Henning</au><au>Patel, Dinshaw</au><au>Meister, Gunter</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phosphorylation of human Argonaute proteins affects small RNA binding</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>2011-03-01</date><risdate>2011</risdate><volume>39</volume><issue>6</issue><spage>2330</spage><epage>2343</epage><pages>2330-2343</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><abstract>Argonaute (Ago) proteins are highly conserved between species and constitute a direct-binding platform for small RNAs including short-interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi interacting RNAs (piRNAs). Small RNAs function as guides whereas Ago proteins are the actual mediators of gene silencing. Although the major steps in RNA-guided gene silencing have been elucidated, not much is known about Ago-protein regulation. Here we report a comprehensive analysis of Ago2 phosphorylation in human cells. We find that the highly conserved tyrosine Y529, located in the small RNA 5'-end-binding pocket of Ago proteins can be phosphorylated. By substituting Y529 with a negatively charged glutamate (E) mimicking a phosphorylated tyrosine, we show that small RNA binding is strongly reduced. Our data suggest that a negatively charged phospho-tyrosine generates a repulsive force that prevents efficient binding of the negatively charged 5' phosphate of the small RNA.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>21071408</pmid><doi>10.1093/nar/gkq1032</doi><tpages>14</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0305-1048 |
| ispartof | Nucleic acids research, 2011-03, Vol.39 (6), p.2330-2343 |
| issn | 0305-1048 1362-4962 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3064767 |
| source | PubMed Central Free; MEDLINE; DOAJ Directory of Open Access Journals; Oxford Journals Open Access Collection; Free Full-Text Journals in Chemistry |
| subjects | Argonaute 2 protein Argonaute Proteins Data processing Eukaryotic Initiation Factor-2 - chemistry Eukaryotic Initiation Factor-2 - genetics Eukaryotic Initiation Factor-2 - metabolism Eukaryotic Initiation Factors - chemistry Eukaryotic Initiation Factors - genetics Eukaryotic Initiation Factors - metabolism Gene silencing Glutamic acid HEK293 Cells Humans Mimicry miRNA Mutation Phosphate Phosphorylation Phosphotyrosine - chemistry Protein Binding Protein Structure, Tertiary Ribonuclease III - metabolism RNA RNA, Small Interfering - metabolism RNA, Small Untranslated - metabolism RNA-Binding Proteins - metabolism siRNA Tyrosine Tyrosine - metabolism |
| title | Phosphorylation of human Argonaute proteins affects small RNA binding |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-20T11%3A02%3A25IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Phosphorylation%20of%20human%20Argonaute%20proteins%20affects%20small%20RNA%20binding&rft.jtitle=Nucleic%20acids%20research&rft.au=R%C3%BCdel,%20Sabine&rft.date=2011-03-01&rft.volume=39&rft.issue=6&rft.spage=2330&rft.epage=2343&rft.pages=2330-2343&rft.issn=0305-1048&rft.eissn=1362-4962&rft_id=info:doi/10.1093/nar/gkq1032&rft_dat=%3Cproquest_pubme%3E899157436%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=865693821&rft_id=info:pmid/21071408&rfr_iscdi=true |