The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping
Eukaryotic mRNAs are modified at their 5′ end early during transcription by the addition of N 7-methylguanosine (m 7 G), which forms the “cap” on the first 5′ nucleotide. Identification of the 5′ nucleotide on mRNA is necessary for determination of the Transcription Start Site (TSS). We explored the...
Gespeichert in:
| Veröffentlicht in: | Scientific reports 2019-06, Vol.9 (1), p.8594-9, Article 8594 |
|---|---|
| Hauptverfasser: | , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 9 |
|---|---|
| container_issue | 1 |
| container_start_page | 8594 |
| container_title | Scientific reports |
| container_volume | 9 |
| creator | Wulf, Madalee G. Buswell, John Chan, Siu-Hong Dai, Nan Marks, Katherine Martin, Evan R. Tzertzinis, George Whipple, Joseph M. Corrêa, Ivan R. Schildkraut, Ira |
| description | Eukaryotic mRNAs are modified at their 5′ end early during transcription by the addition of
N
7-methylguanosine (m
7
G), which forms the “cap” on the first 5′ nucleotide. Identification of the 5′ nucleotide on mRNA is necessary for determination of the Transcription Start Site (TSS). We explored the effect of various reaction conditions on the activity of the yeast scavenger mRNA decapping enzyme DcpS and examined decapping of 30 chemically distinct cap structures varying the state of methylation, sugar, phosphate linkage, and base composition on 25mer RNA oligonucleotides. Contrary to the generally accepted belief that DcpS enzymes only decap short oligonucleotides, we found that the yeast scavenger decapping enzyme decaps RNA transcripts as long as 1400 nucleotides. Further, we validated the application of yDcpS for enriching capped RNA using a strategy of specifically tagging the 5′ end of capped RNA by first decapping and then recapping it with an affinity-tagged guanosine nucleotide. |
| doi_str_mv | 10.1038/s41598-019-45083-5 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6565619</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2239637554</sourcerecordid><originalsourceid>FETCH-LOGICAL-c511t-e5839740e8b19c159e108c0f5befae77f2eae2d8107ce32d29f593923cff85283</originalsourceid><addsrcrecordid>eNp9kUtrGzEUhUVpaILjP5BFEXTTzaR6jDLSpmDcpimEBPKA7ISsuXIUxtJUGhucX185dh7tIpJAgvudc3U5CB1RckwJl99yTYWSFaGqqgWRvBIf0AEjtagYZ-zjm_c-Guf8QMoSTNVUfUL7nFLVlHOA7m7uAa_B5AFna1YQ5pBwC9b0vQ9zDOFxvQD8w_bX2IQW-yHjUuq8NYOPAbuYsA945YcU8dXFBKdn6SHac6bLMN7dI3R7-vNmeladX_76PZ2cV1ZQOlQgJFdNTUDOqLJlJKBEWuLEDJyBpnEMDLBWUtJY4KxlygnFFePWOSmY5CP0fevbL2cLaC2EIZlO98kvTFrraLz-txL8vZ7HlT4RZVNVDL7uDFL8s4Q86IXPFrrOBIjLrBmrC8k2PUfoy3_oQ1ymUMYrFFcnvBGiLhTbUjbFnBO4l89QojfZ6W12umSnn7LToog-vx3jRfKcVAH4FsiltEnptfc7tn8B0FSkyg</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2239637554</pqid></control><display><type>article</type><title>The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping</title><source>MEDLINE</source><source>Nature Free</source><source>DOAJ Directory of Open Access Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>Free Full-Text Journals in Chemistry</source><source>Springer Nature OA Free Journals</source><creator>Wulf, Madalee G. ; Buswell, John ; Chan, Siu-Hong ; Dai, Nan ; Marks, Katherine ; Martin, Evan R. ; Tzertzinis, George ; Whipple, Joseph M. ; Corrêa, Ivan R. ; Schildkraut, Ira</creator><creatorcontrib>Wulf, Madalee G. ; Buswell, John ; Chan, Siu-Hong ; Dai, Nan ; Marks, Katherine ; Martin, Evan R. ; Tzertzinis, George ; Whipple, Joseph M. ; Corrêa, Ivan R. ; Schildkraut, Ira</creatorcontrib><description>Eukaryotic mRNAs are modified at their 5′ end early during transcription by the addition of
N
7-methylguanosine (m
7
G), which forms the “cap” on the first 5′ nucleotide. Identification of the 5′ nucleotide on mRNA is necessary for determination of the Transcription Start Site (TSS). We explored the effect of various reaction conditions on the activity of the yeast scavenger mRNA decapping enzyme DcpS and examined decapping of 30 chemically distinct cap structures varying the state of methylation, sugar, phosphate linkage, and base composition on 25mer RNA oligonucleotides. Contrary to the generally accepted belief that DcpS enzymes only decap short oligonucleotides, we found that the yeast scavenger decapping enzyme decaps RNA transcripts as long as 1400 nucleotides. Further, we validated the application of yDcpS for enriching capped RNA using a strategy of specifically tagging the 5′ end of capped RNA by first decapping and then recapping it with an affinity-tagged guanosine nucleotide.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-019-45083-5</identifier><identifier>PMID: 31197197</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>38/71 ; 38/90 ; 631/337/1645 ; 631/337/572 ; 631/45/500 ; 631/92/500 ; 82/29 ; Base composition ; Diphosphates - metabolism ; Endoribonucleases - metabolism ; Enzymes ; Guanosine ; Humanities and Social Sciences ; Humans ; Hydrogen-Ion Concentration ; Hydrolysis ; multidisciplinary ; Nucleic Acid Conformation ; Nucleotides ; Oligonucleotides ; Osmolar Concentration ; RNA Cap Analogs - metabolism ; RNA Caps - chemistry ; RNA Caps - metabolism ; RNA, Messenger - chemistry ; RNA, Messenger - metabolism ; Saccharomyces cerevisiae - enzymology ; Saccharomyces cerevisiae Proteins - metabolism ; Science ; Science (multidisciplinary) ; Sugar ; Transcription ; Yeast</subject><ispartof>Scientific reports, 2019-06, Vol.9 (1), p.8594-9, Article 8594</ispartof><rights>The Author(s) 2019</rights><rights>2019. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c511t-e5839740e8b19c159e108c0f5befae77f2eae2d8107ce32d29f593923cff85283</citedby><cites>FETCH-LOGICAL-c511t-e5839740e8b19c159e108c0f5befae77f2eae2d8107ce32d29f593923cff85283</cites><orcidid>0000-0002-3169-6878</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6565619/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6565619/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,27901,27902,41096,42165,51551,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31197197$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wulf, Madalee G.</creatorcontrib><creatorcontrib>Buswell, John</creatorcontrib><creatorcontrib>Chan, Siu-Hong</creatorcontrib><creatorcontrib>Dai, Nan</creatorcontrib><creatorcontrib>Marks, Katherine</creatorcontrib><creatorcontrib>Martin, Evan R.</creatorcontrib><creatorcontrib>Tzertzinis, George</creatorcontrib><creatorcontrib>Whipple, Joseph M.</creatorcontrib><creatorcontrib>Corrêa, Ivan R.</creatorcontrib><creatorcontrib>Schildkraut, Ira</creatorcontrib><title>The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Eukaryotic mRNAs are modified at their 5′ end early during transcription by the addition of
N
7-methylguanosine (m
7
G), which forms the “cap” on the first 5′ nucleotide. Identification of the 5′ nucleotide on mRNA is necessary for determination of the Transcription Start Site (TSS). We explored the effect of various reaction conditions on the activity of the yeast scavenger mRNA decapping enzyme DcpS and examined decapping of 30 chemically distinct cap structures varying the state of methylation, sugar, phosphate linkage, and base composition on 25mer RNA oligonucleotides. Contrary to the generally accepted belief that DcpS enzymes only decap short oligonucleotides, we found that the yeast scavenger decapping enzyme decaps RNA transcripts as long as 1400 nucleotides. Further, we validated the application of yDcpS for enriching capped RNA using a strategy of specifically tagging the 5′ end of capped RNA by first decapping and then recapping it with an affinity-tagged guanosine nucleotide.</description><subject>38/71</subject><subject>38/90</subject><subject>631/337/1645</subject><subject>631/337/572</subject><subject>631/45/500</subject><subject>631/92/500</subject><subject>82/29</subject><subject>Base composition</subject><subject>Diphosphates - metabolism</subject><subject>Endoribonucleases - metabolism</subject><subject>Enzymes</subject><subject>Guanosine</subject><subject>Humanities and Social Sciences</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>Hydrolysis</subject><subject>multidisciplinary</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleotides</subject><subject>Oligonucleotides</subject><subject>Osmolar Concentration</subject><subject>RNA Cap Analogs - metabolism</subject><subject>RNA Caps - chemistry</subject><subject>RNA Caps - metabolism</subject><subject>RNA, Messenger - chemistry</subject><subject>RNA, Messenger - metabolism</subject><subject>Saccharomyces cerevisiae - enzymology</subject><subject>Saccharomyces cerevisiae Proteins - metabolism</subject><subject>Science</subject><subject>Science (multidisciplinary)</subject><subject>Sugar</subject><subject>Transcription</subject><subject>Yeast</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9kUtrGzEUhUVpaILjP5BFEXTTzaR6jDLSpmDcpimEBPKA7ISsuXIUxtJUGhucX185dh7tIpJAgvudc3U5CB1RckwJl99yTYWSFaGqqgWRvBIf0AEjtagYZ-zjm_c-Guf8QMoSTNVUfUL7nFLVlHOA7m7uAa_B5AFna1YQ5pBwC9b0vQ9zDOFxvQD8w_bX2IQW-yHjUuq8NYOPAbuYsA945YcU8dXFBKdn6SHac6bLMN7dI3R7-vNmeladX_76PZ2cV1ZQOlQgJFdNTUDOqLJlJKBEWuLEDJyBpnEMDLBWUtJY4KxlygnFFePWOSmY5CP0fevbL2cLaC2EIZlO98kvTFrraLz-txL8vZ7HlT4RZVNVDL7uDFL8s4Q86IXPFrrOBIjLrBmrC8k2PUfoy3_oQ1ymUMYrFFcnvBGiLhTbUjbFnBO4l89QojfZ6W12umSnn7LToog-vx3jRfKcVAH4FsiltEnptfc7tn8B0FSkyg</recordid><startdate>20190613</startdate><enddate>20190613</enddate><creator>Wulf, Madalee G.</creator><creator>Buswell, John</creator><creator>Chan, Siu-Hong</creator><creator>Dai, Nan</creator><creator>Marks, Katherine</creator><creator>Martin, Evan R.</creator><creator>Tzertzinis, George</creator><creator>Whipple, Joseph M.</creator><creator>Corrêa, Ivan R.</creator><creator>Schildkraut, Ira</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</scope><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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-3169-6878</orcidid></search><sort><creationdate>20190613</creationdate><title>The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping</title><author>Wulf, Madalee G. ; Buswell, John ; Chan, Siu-Hong ; Dai, Nan ; Marks, Katherine ; Martin, Evan R. ; Tzertzinis, George ; Whipple, Joseph M. ; Corrêa, Ivan R. ; Schildkraut, Ira</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c511t-e5839740e8b19c159e108c0f5befae77f2eae2d8107ce32d29f593923cff85283</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>38/71</topic><topic>38/90</topic><topic>631/337/1645</topic><topic>631/337/572</topic><topic>631/45/500</topic><topic>631/92/500</topic><topic>82/29</topic><topic>Base composition</topic><topic>Diphosphates - metabolism</topic><topic>Endoribonucleases - metabolism</topic><topic>Enzymes</topic><topic>Guanosine</topic><topic>Humanities and Social Sciences</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>Hydrolysis</topic><topic>multidisciplinary</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleotides</topic><topic>Oligonucleotides</topic><topic>Osmolar Concentration</topic><topic>RNA Cap Analogs - metabolism</topic><topic>RNA Caps - chemistry</topic><topic>RNA Caps - metabolism</topic><topic>RNA, Messenger - chemistry</topic><topic>RNA, Messenger - metabolism</topic><topic>Saccharomyces cerevisiae - enzymology</topic><topic>Saccharomyces cerevisiae Proteins - metabolism</topic><topic>Science</topic><topic>Science (multidisciplinary)</topic><topic>Sugar</topic><topic>Transcription</topic><topic>Yeast</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wulf, Madalee G.</creatorcontrib><creatorcontrib>Buswell, John</creatorcontrib><creatorcontrib>Chan, Siu-Hong</creatorcontrib><creatorcontrib>Dai, Nan</creatorcontrib><creatorcontrib>Marks, Katherine</creatorcontrib><creatorcontrib>Martin, Evan R.</creatorcontrib><creatorcontrib>Tzertzinis, George</creatorcontrib><creatorcontrib>Whipple, Joseph M.</creatorcontrib><creatorcontrib>Corrêa, Ivan R.</creatorcontrib><creatorcontrib>Schildkraut, Ira</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wulf, Madalee G.</au><au>Buswell, John</au><au>Chan, Siu-Hong</au><au>Dai, Nan</au><au>Marks, Katherine</au><au>Martin, Evan R.</au><au>Tzertzinis, George</au><au>Whipple, Joseph M.</au><au>Corrêa, Ivan R.</au><au>Schildkraut, Ira</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2019-06-13</date><risdate>2019</risdate><volume>9</volume><issue>1</issue><spage>8594</spage><epage>9</epage><pages>8594-9</pages><artnum>8594</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Eukaryotic mRNAs are modified at their 5′ end early during transcription by the addition of
N
7-methylguanosine (m
7
G), which forms the “cap” on the first 5′ nucleotide. Identification of the 5′ nucleotide on mRNA is necessary for determination of the Transcription Start Site (TSS). We explored the effect of various reaction conditions on the activity of the yeast scavenger mRNA decapping enzyme DcpS and examined decapping of 30 chemically distinct cap structures varying the state of methylation, sugar, phosphate linkage, and base composition on 25mer RNA oligonucleotides. Contrary to the generally accepted belief that DcpS enzymes only decap short oligonucleotides, we found that the yeast scavenger decapping enzyme decaps RNA transcripts as long as 1400 nucleotides. Further, we validated the application of yDcpS for enriching capped RNA using a strategy of specifically tagging the 5′ end of capped RNA by first decapping and then recapping it with an affinity-tagged guanosine nucleotide.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>31197197</pmid><doi>10.1038/s41598-019-45083-5</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-3169-6878</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 2045-2322 |
| ispartof | Scientific reports, 2019-06, Vol.9 (1), p.8594-9, Article 8594 |
| issn | 2045-2322 2045-2322 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6565619 |
| source | MEDLINE; Nature Free; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry; Springer Nature OA Free Journals |
| subjects | 38/71 38/90 631/337/1645 631/337/572 631/45/500 631/92/500 82/29 Base composition Diphosphates - metabolism Endoribonucleases - metabolism Enzymes Guanosine Humanities and Social Sciences Humans Hydrogen-Ion Concentration Hydrolysis multidisciplinary Nucleic Acid Conformation Nucleotides Oligonucleotides Osmolar Concentration RNA Cap Analogs - metabolism RNA Caps - chemistry RNA Caps - metabolism RNA, Messenger - chemistry RNA, Messenger - metabolism Saccharomyces cerevisiae - enzymology Saccharomyces cerevisiae Proteins - metabolism Science Science (multidisciplinary) Sugar Transcription Yeast |
| title | The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-11T11%3A41%3A29IST&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=The%20yeast%20scavenger%20decapping%20enzyme%20DcpS%20and%20its%20application%20for%20in%20vitro%20RNA%20recapping&rft.jtitle=Scientific%20reports&rft.au=Wulf,%20Madalee%20G.&rft.date=2019-06-13&rft.volume=9&rft.issue=1&rft.spage=8594&rft.epage=9&rft.pages=8594-9&rft.artnum=8594&rft.issn=2045-2322&rft.eissn=2045-2322&rft_id=info:doi/10.1038/s41598-019-45083-5&rft_dat=%3Cproquest_pubme%3E2239637554%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=2239637554&rft_id=info:pmid/31197197&rfr_iscdi=true |