Loading…

The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation

PML nuclear bodies (NBs) are accumulations of cellular proteins embedded in a scaffold-like structure built by SUMO-modified PML/TRIM19. PML and other NB proteins act as cellular restriction factors against human cytomegalovirus (HCMV); however, this intrinsic defense is counteracted by the immediat...

Full description

Saved in:
Bibliographic Details
Published in:Journal of virology 2017-02, Vol.91 (4)
Main Authors: Schilling, Eva-Maria, Scherer, Myriam, Reuter, Nina, Schweininger, Johannes, Muller, Yves A, Stamminger, Thomas
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-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23
cites cdi_FETCH-LOGICAL-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23
container_end_page
container_issue 4
container_start_page
container_title Journal of virology
container_volume 91
creator Schilling, Eva-Maria
Scherer, Myriam
Reuter, Nina
Schweininger, Johannes
Muller, Yves A
Stamminger, Thomas
description PML nuclear bodies (NBs) are accumulations of cellular proteins embedded in a scaffold-like structure built by SUMO-modified PML/TRIM19. PML and other NB proteins act as cellular restriction factors against human cytomegalovirus (HCMV); however, this intrinsic defense is counteracted by the immediate early protein 1 (IE1) of HCMV. IE1 directly interacts with the PML coiled-coil domain via its globular core region and disrupts NB foci by inducing a loss of PML SUMOylation. Here, we demonstrate that IE1 acts via abrogating the de novo SUMOylation of PML. In order to overcome reversible SUMOylation dynamics, we made use of a cell-based assay that combines inducible IE1 expression with a SUMO mutant resistant to SUMO proteases. Interestingly, we observed that IE1 expression did not affect preSUMOylated PML; however, it clearly prevented de novo SUMO conjugation. Consistent results were obtained by in vitro SUMOylation assays, demonstrating that IE1 alone is sufficient for this effect. Furthermore, IE1 acts in a selective manner, since K160 was identified as the main target lysine. This is strengthened by the fact that IE1 also prevents As O -mediated hyperSUMOylation of K160, thereby blocking PML degradation. Since IE1 did not interfere with coiled-coil-mediated PML dimerization, we propose that IE1 affects PML autoSUMOylation either by directly abrogating PML E3 ligase function or by preventing access to SUMO sites. Thus, our data suggest a novel mechanism for how a viral protein counteracts a cellular restriction factor by selectively preventing the de novo SUMOylation at specific lysine residues without affecting global protein SUMOylation. The human cytomegalovirus IE1 protein acts as an important antagonist of a cellular restriction mechanism that is mediated by subnuclear structures termed PML nuclear bodies. This function of IE1 is required for efficient viral replication and thus constitutes a potential target for antiviral strategies. In this paper, we further elucidate the molecular mechanism for how IE1 antagonizes PML NBs. We show that tight binding of IE1 to PML interferes with the de novo SUMOylation of a distinct lysine residue that is also the target of stress-mediated hyperSUMOylation of PML. This is of importance since it represents a novel mechanism used by a viral antagonist of intrinsic immunity. Furthermore, it highlights the possibility of developing small molecules that specifically abrogate this PML-antagonistic activity of IE1 and thu
doi_str_mv 10.1128/JVI.02049-16
format article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5286881</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1845252247</sourcerecordid><originalsourceid>FETCH-LOGICAL-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23</originalsourceid><addsrcrecordid>eNqNkU1v1DAYhC0Eokvhxhn5yKEp_sw6F6SyLW2q3bYSLeJmOY6za5TYxXZWCr-BH11vvwQ3Tu9hHs3MqwHgPUaHGBPx6fx7fYgIYlWByxdghlElCs4xewlmCBFScCp-7IE3Mf5ECDNWstdgj8wrRAWiM_DnemPg2TgoBxdT8oNZq95vbRgjrE8wvAo-GevgkUtq7Z39bSK8Wi3hxah7owL84tupWJnWqmRaWLsUrItWw3oYRmfTBLdWwZQjarexjU3WO-i7e4tjAy_81sNvN6vLqVc76S141ak-mnePdx_cfD25XpwVy8vTenG0LDQrUSoqLqoKaUJEyTjFTaMo7TjBWnUdF8pg3RHBDObNHOOOcdK2QuuKNx1VWSd0H3x-8L0dm8G02uTeqpe3wQ4qTNIrK_9VnN3Itd9KniOFwNng46NB8L9GE5McbNSm75UzfowSZ4xSVIr5f6C5ICeE7dCDB1QHH2Mw3XMjjORua5m3lvdbS1xm_MPfXzzDT-PSO6aTpdc</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1845252247</pqid></control><display><type>article</type><title>The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation</title><source>American Society for Microbiology</source><source>PubMed Central</source><creator>Schilling, Eva-Maria ; Scherer, Myriam ; Reuter, Nina ; Schweininger, Johannes ; Muller, Yves A ; Stamminger, Thomas</creator><contributor>Sandri-Goldin, Rozanne M.</contributor><creatorcontrib>Schilling, Eva-Maria ; Scherer, Myriam ; Reuter, Nina ; Schweininger, Johannes ; Muller, Yves A ; Stamminger, Thomas ; Sandri-Goldin, Rozanne M.</creatorcontrib><description>PML nuclear bodies (NBs) are accumulations of cellular proteins embedded in a scaffold-like structure built by SUMO-modified PML/TRIM19. PML and other NB proteins act as cellular restriction factors against human cytomegalovirus (HCMV); however, this intrinsic defense is counteracted by the immediate early protein 1 (IE1) of HCMV. IE1 directly interacts with the PML coiled-coil domain via its globular core region and disrupts NB foci by inducing a loss of PML SUMOylation. Here, we demonstrate that IE1 acts via abrogating the de novo SUMOylation of PML. In order to overcome reversible SUMOylation dynamics, we made use of a cell-based assay that combines inducible IE1 expression with a SUMO mutant resistant to SUMO proteases. Interestingly, we observed that IE1 expression did not affect preSUMOylated PML; however, it clearly prevented de novo SUMO conjugation. Consistent results were obtained by in vitro SUMOylation assays, demonstrating that IE1 alone is sufficient for this effect. Furthermore, IE1 acts in a selective manner, since K160 was identified as the main target lysine. This is strengthened by the fact that IE1 also prevents As O -mediated hyperSUMOylation of K160, thereby blocking PML degradation. Since IE1 did not interfere with coiled-coil-mediated PML dimerization, we propose that IE1 affects PML autoSUMOylation either by directly abrogating PML E3 ligase function or by preventing access to SUMO sites. Thus, our data suggest a novel mechanism for how a viral protein counteracts a cellular restriction factor by selectively preventing the de novo SUMOylation at specific lysine residues without affecting global protein SUMOylation. The human cytomegalovirus IE1 protein acts as an important antagonist of a cellular restriction mechanism that is mediated by subnuclear structures termed PML nuclear bodies. This function of IE1 is required for efficient viral replication and thus constitutes a potential target for antiviral strategies. In this paper, we further elucidate the molecular mechanism for how IE1 antagonizes PML NBs. We show that tight binding of IE1 to PML interferes with the de novo SUMOylation of a distinct lysine residue that is also the target of stress-mediated hyperSUMOylation of PML. This is of importance since it represents a novel mechanism used by a viral antagonist of intrinsic immunity. Furthermore, it highlights the possibility of developing small molecules that specifically abrogate this PML-antagonistic activity of IE1 and thus inhibit viral replication.</description><identifier>ISSN: 0022-538X</identifier><identifier>EISSN: 1098-5514</identifier><identifier>DOI: 10.1128/JVI.02049-16</identifier><identifier>PMID: 27903803</identifier><language>eng</language><publisher>United States: American Society for Microbiology</publisher><subject>Cell Line ; Cytomegalovirus - physiology ; Cytomegalovirus Infections - immunology ; Cytomegalovirus Infections - metabolism ; Cytomegalovirus Infections - virology ; Enzyme Stability ; Herpesviridae ; Human cytomegalovirus ; Humans ; Immediate-Early Proteins - metabolism ; Immunity ; Intranuclear Inclusion Bodies - metabolism ; Mutation ; Promyelocytic Leukemia Protein - metabolism ; Small Ubiquitin-Related Modifier Proteins - genetics ; Small Ubiquitin-Related Modifier Proteins - metabolism ; Sumoylation ; Virus-Cell Interactions</subject><ispartof>Journal of virology, 2017-02, Vol.91 (4)</ispartof><rights>Copyright © 2017 American Society for Microbiology.</rights><rights>Copyright © 2017 American Society for Microbiology. 2017 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23</citedby><cites>FETCH-LOGICAL-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23</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/PMC5286881/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5286881/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27903803$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Sandri-Goldin, Rozanne M.</contributor><creatorcontrib>Schilling, Eva-Maria</creatorcontrib><creatorcontrib>Scherer, Myriam</creatorcontrib><creatorcontrib>Reuter, Nina</creatorcontrib><creatorcontrib>Schweininger, Johannes</creatorcontrib><creatorcontrib>Muller, Yves A</creatorcontrib><creatorcontrib>Stamminger, Thomas</creatorcontrib><title>The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation</title><title>Journal of virology</title><addtitle>J Virol</addtitle><description>PML nuclear bodies (NBs) are accumulations of cellular proteins embedded in a scaffold-like structure built by SUMO-modified PML/TRIM19. PML and other NB proteins act as cellular restriction factors against human cytomegalovirus (HCMV); however, this intrinsic defense is counteracted by the immediate early protein 1 (IE1) of HCMV. IE1 directly interacts with the PML coiled-coil domain via its globular core region and disrupts NB foci by inducing a loss of PML SUMOylation. Here, we demonstrate that IE1 acts via abrogating the de novo SUMOylation of PML. In order to overcome reversible SUMOylation dynamics, we made use of a cell-based assay that combines inducible IE1 expression with a SUMO mutant resistant to SUMO proteases. Interestingly, we observed that IE1 expression did not affect preSUMOylated PML; however, it clearly prevented de novo SUMO conjugation. Consistent results were obtained by in vitro SUMOylation assays, demonstrating that IE1 alone is sufficient for this effect. Furthermore, IE1 acts in a selective manner, since K160 was identified as the main target lysine. This is strengthened by the fact that IE1 also prevents As O -mediated hyperSUMOylation of K160, thereby blocking PML degradation. Since IE1 did not interfere with coiled-coil-mediated PML dimerization, we propose that IE1 affects PML autoSUMOylation either by directly abrogating PML E3 ligase function or by preventing access to SUMO sites. Thus, our data suggest a novel mechanism for how a viral protein counteracts a cellular restriction factor by selectively preventing the de novo SUMOylation at specific lysine residues without affecting global protein SUMOylation. The human cytomegalovirus IE1 protein acts as an important antagonist of a cellular restriction mechanism that is mediated by subnuclear structures termed PML nuclear bodies. This function of IE1 is required for efficient viral replication and thus constitutes a potential target for antiviral strategies. In this paper, we further elucidate the molecular mechanism for how IE1 antagonizes PML NBs. We show that tight binding of IE1 to PML interferes with the de novo SUMOylation of a distinct lysine residue that is also the target of stress-mediated hyperSUMOylation of PML. This is of importance since it represents a novel mechanism used by a viral antagonist of intrinsic immunity. Furthermore, it highlights the possibility of developing small molecules that specifically abrogate this PML-antagonistic activity of IE1 and thus inhibit viral replication.</description><subject>Cell Line</subject><subject>Cytomegalovirus - physiology</subject><subject>Cytomegalovirus Infections - immunology</subject><subject>Cytomegalovirus Infections - metabolism</subject><subject>Cytomegalovirus Infections - virology</subject><subject>Enzyme Stability</subject><subject>Herpesviridae</subject><subject>Human cytomegalovirus</subject><subject>Humans</subject><subject>Immediate-Early Proteins - metabolism</subject><subject>Immunity</subject><subject>Intranuclear Inclusion Bodies - metabolism</subject><subject>Mutation</subject><subject>Promyelocytic Leukemia Protein - metabolism</subject><subject>Small Ubiquitin-Related Modifier Proteins - genetics</subject><subject>Small Ubiquitin-Related Modifier Proteins - metabolism</subject><subject>Sumoylation</subject><subject>Virus-Cell Interactions</subject><issn>0022-538X</issn><issn>1098-5514</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><recordid>eNqNkU1v1DAYhC0Eokvhxhn5yKEp_sw6F6SyLW2q3bYSLeJmOY6za5TYxXZWCr-BH11vvwQ3Tu9hHs3MqwHgPUaHGBPx6fx7fYgIYlWByxdghlElCs4xewlmCBFScCp-7IE3Mf5ECDNWstdgj8wrRAWiM_DnemPg2TgoBxdT8oNZq95vbRgjrE8wvAo-GevgkUtq7Z39bSK8Wi3hxah7owL84tupWJnWqmRaWLsUrItWw3oYRmfTBLdWwZQjarexjU3WO-i7e4tjAy_81sNvN6vLqVc76S141ak-mnePdx_cfD25XpwVy8vTenG0LDQrUSoqLqoKaUJEyTjFTaMo7TjBWnUdF8pg3RHBDObNHOOOcdK2QuuKNx1VWSd0H3x-8L0dm8G02uTeqpe3wQ4qTNIrK_9VnN3Itd9KniOFwNng46NB8L9GE5McbNSm75UzfowSZ4xSVIr5f6C5ICeE7dCDB1QHH2Mw3XMjjORua5m3lvdbS1xm_MPfXzzDT-PSO6aTpdc</recordid><startdate>20170215</startdate><enddate>20170215</enddate><creator>Schilling, Eva-Maria</creator><creator>Scherer, Myriam</creator><creator>Reuter, Nina</creator><creator>Schweininger, Johannes</creator><creator>Muller, Yves A</creator><creator>Stamminger, Thomas</creator><general>American Society for Microbiology</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>7T5</scope><scope>7U9</scope><scope>H94</scope><scope>5PM</scope></search><sort><creationdate>20170215</creationdate><title>The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation</title><author>Schilling, Eva-Maria ; Scherer, Myriam ; Reuter, Nina ; Schweininger, Johannes ; Muller, Yves A ; Stamminger, Thomas</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Cell Line</topic><topic>Cytomegalovirus - physiology</topic><topic>Cytomegalovirus Infections - immunology</topic><topic>Cytomegalovirus Infections - metabolism</topic><topic>Cytomegalovirus Infections - virology</topic><topic>Enzyme Stability</topic><topic>Herpesviridae</topic><topic>Human cytomegalovirus</topic><topic>Humans</topic><topic>Immediate-Early Proteins - metabolism</topic><topic>Immunity</topic><topic>Intranuclear Inclusion Bodies - metabolism</topic><topic>Mutation</topic><topic>Promyelocytic Leukemia Protein - metabolism</topic><topic>Small Ubiquitin-Related Modifier Proteins - genetics</topic><topic>Small Ubiquitin-Related Modifier Proteins - metabolism</topic><topic>Sumoylation</topic><topic>Virus-Cell Interactions</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schilling, Eva-Maria</creatorcontrib><creatorcontrib>Scherer, Myriam</creatorcontrib><creatorcontrib>Reuter, Nina</creatorcontrib><creatorcontrib>Schweininger, Johannes</creatorcontrib><creatorcontrib>Muller, Yves A</creatorcontrib><creatorcontrib>Stamminger, Thomas</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>Immunology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schilling, Eva-Maria</au><au>Scherer, Myriam</au><au>Reuter, Nina</au><au>Schweininger, Johannes</au><au>Muller, Yves A</au><au>Stamminger, Thomas</au><au>Sandri-Goldin, Rozanne M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation</atitle><jtitle>Journal of virology</jtitle><addtitle>J Virol</addtitle><date>2017-02-15</date><risdate>2017</risdate><volume>91</volume><issue>4</issue><issn>0022-538X</issn><eissn>1098-5514</eissn><abstract>PML nuclear bodies (NBs) are accumulations of cellular proteins embedded in a scaffold-like structure built by SUMO-modified PML/TRIM19. PML and other NB proteins act as cellular restriction factors against human cytomegalovirus (HCMV); however, this intrinsic defense is counteracted by the immediate early protein 1 (IE1) of HCMV. IE1 directly interacts with the PML coiled-coil domain via its globular core region and disrupts NB foci by inducing a loss of PML SUMOylation. Here, we demonstrate that IE1 acts via abrogating the de novo SUMOylation of PML. In order to overcome reversible SUMOylation dynamics, we made use of a cell-based assay that combines inducible IE1 expression with a SUMO mutant resistant to SUMO proteases. Interestingly, we observed that IE1 expression did not affect preSUMOylated PML; however, it clearly prevented de novo SUMO conjugation. Consistent results were obtained by in vitro SUMOylation assays, demonstrating that IE1 alone is sufficient for this effect. Furthermore, IE1 acts in a selective manner, since K160 was identified as the main target lysine. This is strengthened by the fact that IE1 also prevents As O -mediated hyperSUMOylation of K160, thereby blocking PML degradation. Since IE1 did not interfere with coiled-coil-mediated PML dimerization, we propose that IE1 affects PML autoSUMOylation either by directly abrogating PML E3 ligase function or by preventing access to SUMO sites. Thus, our data suggest a novel mechanism for how a viral protein counteracts a cellular restriction factor by selectively preventing the de novo SUMOylation at specific lysine residues without affecting global protein SUMOylation. The human cytomegalovirus IE1 protein acts as an important antagonist of a cellular restriction mechanism that is mediated by subnuclear structures termed PML nuclear bodies. This function of IE1 is required for efficient viral replication and thus constitutes a potential target for antiviral strategies. In this paper, we further elucidate the molecular mechanism for how IE1 antagonizes PML NBs. We show that tight binding of IE1 to PML interferes with the de novo SUMOylation of a distinct lysine residue that is also the target of stress-mediated hyperSUMOylation of PML. This is of importance since it represents a novel mechanism used by a viral antagonist of intrinsic immunity. Furthermore, it highlights the possibility of developing small molecules that specifically abrogate this PML-antagonistic activity of IE1 and thus inhibit viral replication.</abstract><cop>United States</cop><pub>American Society for Microbiology</pub><pmid>27903803</pmid><doi>10.1128/JVI.02049-16</doi><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0022-538X
ispartof Journal of virology, 2017-02, Vol.91 (4)
issn 0022-538X
1098-5514
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5286881
source American Society for Microbiology; PubMed Central
subjects Cell Line
Cytomegalovirus - physiology
Cytomegalovirus Infections - immunology
Cytomegalovirus Infections - metabolism
Cytomegalovirus Infections - virology
Enzyme Stability
Herpesviridae
Human cytomegalovirus
Humans
Immediate-Early Proteins - metabolism
Immunity
Intranuclear Inclusion Bodies - metabolism
Mutation
Promyelocytic Leukemia Protein - metabolism
Small Ubiquitin-Related Modifier Proteins - genetics
Small Ubiquitin-Related Modifier Proteins - metabolism
Sumoylation
Virus-Cell Interactions
title The Human Cytomegalovirus IE1 Protein Antagonizes PML Nuclear Body-Mediated Intrinsic Immunity via the Inhibition of PML De Novo SUMOylation
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T02%3A47%3A48IST&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%20Human%20Cytomegalovirus%20IE1%20Protein%20Antagonizes%20PML%20Nuclear%20Body-Mediated%20Intrinsic%20Immunity%20via%20the%20Inhibition%20of%20PML%20De%20Novo%20SUMOylation&rft.jtitle=Journal%20of%20virology&rft.au=Schilling,%20Eva-Maria&rft.date=2017-02-15&rft.volume=91&rft.issue=4&rft.issn=0022-538X&rft.eissn=1098-5514&rft_id=info:doi/10.1128/JVI.02049-16&rft_dat=%3Cproquest_pubme%3E1845252247%3C/proquest_pubme%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c460t-958990c22864531bba33f521caff58ae1cf284e15b711f452dd8cc95bf3a58a23%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1845252247&rft_id=info:pmid/27903803&rfr_iscdi=true