Loading…
The biochemical basis for the cooperative action of microRNAs
In cells, closely spaced microRNA (miRNA) target sites within a messenger RNA (mRNA) can act cooperatively, leading to more repression of the target mRNA than expected by independent action at each site. Using purified miRNA-Argonaute (AGO2) complexes, synthetic target RNAs, and a purified domain of...
Saved in:
Published in: | Proceedings of the National Academy of Sciences - PNAS 2020-07, Vol.117 (30), p.17764-17774 |
---|---|
Main Authors: | , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
cited_by | cdi_FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173 |
---|---|
cites | cdi_FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173 |
container_end_page | 17774 |
container_issue | 30 |
container_start_page | 17764 |
container_title | Proceedings of the National Academy of Sciences - PNAS |
container_volume | 117 |
creator | Briskin, Daniel Wang, Peter Y. Bartel, David P. |
description | In cells, closely spaced microRNA (miRNA) target sites within a messenger RNA (mRNA) can act cooperatively, leading to more repression of the target mRNA than expected by independent action at each site. Using purified miRNA-Argonaute (AGO2) complexes, synthetic target RNAs, and a purified domain of TNRC6B (GW182 in flies) that is able to simultaneously bind multiple AGO proteins, we examined both the occupancies and binding affinities of miRNA-AGO2 complexes and target RNAs with either one site or two cooperatively spaced sites. On their own, miRNA-AGO2 complexes displayed little if any cooperative binding to dual sites. In contrast, in the presence of the AGO-binding region of TNRC6B, we observed strong cooperative binding to dual sites, with almost no singly bound target RNAs and substantially increased binding affinities and Hill coefficients. Cooperative binding was retained when the two sites were for two different miRNAs or when the two sites were bound to miRNAs loaded into two different AGO paralogs, AGO1 and AGO2. The improved binding affinity was attributable primarily to a reduced rate of dissociation between miRNA-AGO complexes and their dual-site targets. Thus, the multivalent binding of TNRC6 enables cooperative binding of miRNAAGO complexes to target RNAs, thereby explaining the basis of cooperative action. |
doi_str_mv | 10.1073/pnas.1920404117 |
format | article |
fullrecord | <record><control><sourceid>jstor_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7395462</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><jstor_id>26935505</jstor_id><sourcerecordid>26935505</sourcerecordid><originalsourceid>FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173</originalsourceid><addsrcrecordid>eNpdkc1LHEEQxZuQoOvHOSfDgBcvo9Xf04cIIjEGJILouenpqcn2Mju96Z4V8t-nl9WN5lSH96tH1XuEfKZwTkHzi9Xo8jk1DAQISvUHMqNgaK2EgY9kBsB03Qgm9slBzgsAMLKBPbLPmVKUKjYjXx_nWLUh-jkug3dD1bocctXHVE1F8TGuMLkpPGPl_BTiWMW-KmSKDz-v8hH51Lsh4_HLPCRPN98er2_ru_vvP66v7movwUx1yyU3XeNb7IRE77lrUHcKe01bQCOx8ShY3xmgnWJeiVYDCuB9qyUXVPNDcrn1Xa3bJXYexym5wa5SWLr0x0YX7HtlDHP7Kz5bzY0UihWDsxeDFH-vMU92GbLHYXAjxnW2rIRkTAnRFPT0P3QR12ks722oRiqgRhXqYkuVJHJO2O-OoWA31dhNNfZfNWXjy9sfdvxrFwU42QKLPMW005kyXEqQ_C8XS5P8</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2428560196</pqid></control><display><type>article</type><title>The biochemical basis for the cooperative action of microRNAs</title><source>PubMed (Medline)</source><source>JSTOR Archival Journals and Primary Sources Collection</source><creator>Briskin, Daniel ; Wang, Peter Y. ; Bartel, David P.</creator><creatorcontrib>Briskin, Daniel ; Wang, Peter Y. ; Bartel, David P.</creatorcontrib><description>In cells, closely spaced microRNA (miRNA) target sites within a messenger RNA (mRNA) can act cooperatively, leading to more repression of the target mRNA than expected by independent action at each site. Using purified miRNA-Argonaute (AGO2) complexes, synthetic target RNAs, and a purified domain of TNRC6B (GW182 in flies) that is able to simultaneously bind multiple AGO proteins, we examined both the occupancies and binding affinities of miRNA-AGO2 complexes and target RNAs with either one site or two cooperatively spaced sites. On their own, miRNA-AGO2 complexes displayed little if any cooperative binding to dual sites. In contrast, in the presence of the AGO-binding region of TNRC6B, we observed strong cooperative binding to dual sites, with almost no singly bound target RNAs and substantially increased binding affinities and Hill coefficients. Cooperative binding was retained when the two sites were for two different miRNAs or when the two sites were bound to miRNAs loaded into two different AGO paralogs, AGO1 and AGO2. The improved binding affinity was attributable primarily to a reduced rate of dissociation between miRNA-AGO complexes and their dual-site targets. Thus, the multivalent binding of TNRC6 enables cooperative binding of miRNAAGO complexes to target RNAs, thereby explaining the basis of cooperative action.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.1920404117</identifier><identifier>PMID: 32661162</identifier><language>eng</language><publisher>United States: National Academy of Sciences</publisher><subject>Affinity ; Argonaute 2 protein ; Argonaute Proteins - metabolism ; Binding ; Binding Sites ; Biological Sciences ; Gene Expression Regulation ; Gene Silencing ; Humans ; Kinetics ; MicroRNAs ; MicroRNAs - genetics ; miRNA ; Models, Biological ; mRNA ; Protein Binding ; Ribonucleic acid ; RNA ; RNA Interference ; RNA-Binding Proteins - metabolism ; RNA-Induced Silencing Complex - metabolism</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 2020-07, Vol.117 (30), p.17764-17774</ispartof><rights>Copyright National Academy of Sciences Jul 28, 2020</rights><rights>2020</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173</citedby><cites>FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173</cites><orcidid>0000-0002-6129-2815 ; 0000-0002-3872-2856 ; 0000-0002-8331-9257</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/26935505$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/26935505$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793,58238,58471</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32661162$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Briskin, Daniel</creatorcontrib><creatorcontrib>Wang, Peter Y.</creatorcontrib><creatorcontrib>Bartel, David P.</creatorcontrib><title>The biochemical basis for the cooperative action of microRNAs</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>In cells, closely spaced microRNA (miRNA) target sites within a messenger RNA (mRNA) can act cooperatively, leading to more repression of the target mRNA than expected by independent action at each site. Using purified miRNA-Argonaute (AGO2) complexes, synthetic target RNAs, and a purified domain of TNRC6B (GW182 in flies) that is able to simultaneously bind multiple AGO proteins, we examined both the occupancies and binding affinities of miRNA-AGO2 complexes and target RNAs with either one site or two cooperatively spaced sites. On their own, miRNA-AGO2 complexes displayed little if any cooperative binding to dual sites. In contrast, in the presence of the AGO-binding region of TNRC6B, we observed strong cooperative binding to dual sites, with almost no singly bound target RNAs and substantially increased binding affinities and Hill coefficients. Cooperative binding was retained when the two sites were for two different miRNAs or when the two sites were bound to miRNAs loaded into two different AGO paralogs, AGO1 and AGO2. The improved binding affinity was attributable primarily to a reduced rate of dissociation between miRNA-AGO complexes and their dual-site targets. Thus, the multivalent binding of TNRC6 enables cooperative binding of miRNAAGO complexes to target RNAs, thereby explaining the basis of cooperative action.</description><subject>Affinity</subject><subject>Argonaute 2 protein</subject><subject>Argonaute Proteins - metabolism</subject><subject>Binding</subject><subject>Binding Sites</subject><subject>Biological Sciences</subject><subject>Gene Expression Regulation</subject><subject>Gene Silencing</subject><subject>Humans</subject><subject>Kinetics</subject><subject>MicroRNAs</subject><subject>MicroRNAs - genetics</subject><subject>miRNA</subject><subject>Models, Biological</subject><subject>mRNA</subject><subject>Protein Binding</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA Interference</subject><subject>RNA-Binding Proteins - metabolism</subject><subject>RNA-Induced Silencing Complex - metabolism</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNpdkc1LHEEQxZuQoOvHOSfDgBcvo9Xf04cIIjEGJILouenpqcn2Mju96Z4V8t-nl9WN5lSH96tH1XuEfKZwTkHzi9Xo8jk1DAQISvUHMqNgaK2EgY9kBsB03Qgm9slBzgsAMLKBPbLPmVKUKjYjXx_nWLUh-jkug3dD1bocctXHVE1F8TGuMLkpPGPl_BTiWMW-KmSKDz-v8hH51Lsh4_HLPCRPN98er2_ru_vvP66v7movwUx1yyU3XeNb7IRE77lrUHcKe01bQCOx8ShY3xmgnWJeiVYDCuB9qyUXVPNDcrn1Xa3bJXYexym5wa5SWLr0x0YX7HtlDHP7Kz5bzY0UihWDsxeDFH-vMU92GbLHYXAjxnW2rIRkTAnRFPT0P3QR12ks722oRiqgRhXqYkuVJHJO2O-OoWA31dhNNfZfNWXjy9sfdvxrFwU42QKLPMW005kyXEqQ_C8XS5P8</recordid><startdate>20200728</startdate><enddate>20200728</enddate><creator>Briskin, Daniel</creator><creator>Wang, Peter Y.</creator><creator>Bartel, David P.</creator><general>National Academy of Sciences</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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-6129-2815</orcidid><orcidid>https://orcid.org/0000-0002-3872-2856</orcidid><orcidid>https://orcid.org/0000-0002-8331-9257</orcidid></search><sort><creationdate>20200728</creationdate><title>The biochemical basis for the cooperative action of microRNAs</title><author>Briskin, Daniel ; Wang, Peter Y. ; Bartel, David P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Affinity</topic><topic>Argonaute 2 protein</topic><topic>Argonaute Proteins - metabolism</topic><topic>Binding</topic><topic>Binding Sites</topic><topic>Biological Sciences</topic><topic>Gene Expression Regulation</topic><topic>Gene Silencing</topic><topic>Humans</topic><topic>Kinetics</topic><topic>MicroRNAs</topic><topic>MicroRNAs - genetics</topic><topic>miRNA</topic><topic>Models, Biological</topic><topic>mRNA</topic><topic>Protein Binding</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA Interference</topic><topic>RNA-Binding Proteins - metabolism</topic><topic>RNA-Induced Silencing Complex - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Briskin, Daniel</creatorcontrib><creatorcontrib>Wang, Peter Y.</creatorcontrib><creatorcontrib>Bartel, David P.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Briskin, Daniel</au><au>Wang, Peter Y.</au><au>Bartel, David P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The biochemical basis for the cooperative action of microRNAs</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>2020-07-28</date><risdate>2020</risdate><volume>117</volume><issue>30</issue><spage>17764</spage><epage>17774</epage><pages>17764-17774</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>In cells, closely spaced microRNA (miRNA) target sites within a messenger RNA (mRNA) can act cooperatively, leading to more repression of the target mRNA than expected by independent action at each site. Using purified miRNA-Argonaute (AGO2) complexes, synthetic target RNAs, and a purified domain of TNRC6B (GW182 in flies) that is able to simultaneously bind multiple AGO proteins, we examined both the occupancies and binding affinities of miRNA-AGO2 complexes and target RNAs with either one site or two cooperatively spaced sites. On their own, miRNA-AGO2 complexes displayed little if any cooperative binding to dual sites. In contrast, in the presence of the AGO-binding region of TNRC6B, we observed strong cooperative binding to dual sites, with almost no singly bound target RNAs and substantially increased binding affinities and Hill coefficients. Cooperative binding was retained when the two sites were for two different miRNAs or when the two sites were bound to miRNAs loaded into two different AGO paralogs, AGO1 and AGO2. The improved binding affinity was attributable primarily to a reduced rate of dissociation between miRNA-AGO complexes and their dual-site targets. Thus, the multivalent binding of TNRC6 enables cooperative binding of miRNAAGO complexes to target RNAs, thereby explaining the basis of cooperative action.</abstract><cop>United States</cop><pub>National Academy of Sciences</pub><pmid>32661162</pmid><doi>10.1073/pnas.1920404117</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0002-6129-2815</orcidid><orcidid>https://orcid.org/0000-0002-3872-2856</orcidid><orcidid>https://orcid.org/0000-0002-8331-9257</orcidid><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0027-8424 |
ispartof | Proceedings of the National Academy of Sciences - PNAS, 2020-07, Vol.117 (30), p.17764-17774 |
issn | 0027-8424 1091-6490 |
language | eng |
recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7395462 |
source | PubMed (Medline); JSTOR Archival Journals and Primary Sources Collection |
subjects | Affinity Argonaute 2 protein Argonaute Proteins - metabolism Binding Binding Sites Biological Sciences Gene Expression Regulation Gene Silencing Humans Kinetics MicroRNAs MicroRNAs - genetics miRNA Models, Biological mRNA Protein Binding Ribonucleic acid RNA RNA Interference RNA-Binding Proteins - metabolism RNA-Induced Silencing Complex - metabolism |
title | The biochemical basis for the cooperative action of microRNAs |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T20%3A24%3A00IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-jstor_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=The%20biochemical%20basis%20for%20the%20cooperative%20action%20of%20microRNAs&rft.jtitle=Proceedings%20of%20the%20National%20Academy%20of%20Sciences%20-%20PNAS&rft.au=Briskin,%20Daniel&rft.date=2020-07-28&rft.volume=117&rft.issue=30&rft.spage=17764&rft.epage=17774&rft.pages=17764-17774&rft.issn=0027-8424&rft.eissn=1091-6490&rft_id=info:doi/10.1073/pnas.1920404117&rft_dat=%3Cjstor_pubme%3E26935505%3C/jstor_pubme%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c509t-b3539d8cbed45ecc3a8e7d6ef71b0e95e8ce42fd901d62c64b70e403fb7534173%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=2428560196&rft_id=info:pmid/32661162&rft_jstor_id=26935505&rfr_iscdi=true |