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The C Protein Is Recruited to Measles Virus Ribonucleocapsids by the Phosphoprotein
Measles virus (MeV), like all viruses of the order , utilizes a complex consisting of genomic RNA, nucleoprotein, the RNA-dependent RNA polymerase, and a polymerase cofactor, the phosphoprotein (P), for transcription and replication. We previously showed that a recombinant MeV that does not express...
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Published in: | Journal of virology 2020-01, Vol.94 (4) |
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creator | Pfaller, Christian K Bloyet, Louis-Marie Donohue, Ryan C Huff, Amanda L Bartemes, William P Yousaf, Iris Urzua, Erica Clavière, Mathieu Zachary, Marie de Masson d'Autume, Valentin Carson, Sandra Schieferecke, Adam J Meyer, Alyssa J Gerlier, Denis Cattaneo, Roberto |
description | Measles virus (MeV), like all viruses of the order
, utilizes a complex consisting of genomic RNA, nucleoprotein, the RNA-dependent RNA polymerase, and a polymerase cofactor, the phosphoprotein (P), for transcription and replication. We previously showed that a recombinant MeV that does not express another viral protein, C, has severe transcription and replication deficiencies, including a steeper transcription gradient than the parental virus and generation of defective interfering RNA. This virus is attenuated
and
However, how the C protein operates and whether it is a component of the replication complex remained unclear. Here, we show that C associates with the ribonucleocapsid and forms a complex that can be purified by immunoprecipitation or ultracentrifugation. In the presence of detergent, the C protein is retained on purified ribonucleocapsids less efficiently than the P protein and the polymerase. The C protein is recruited to the ribonucleocapsid through its interaction with the P protein, as shown by immunofluorescence microscopy of cells expressing different combinations of viral proteins and by split luciferase complementation assays. Forty amino-terminal C protein residues are dispensable for the interaction with P, and the carboxyl-terminal half of P is sufficient for the interaction with C. Thus, the C protein, rather than being an "accessory" protein as qualified in textbooks so far, is a ribonucleocapsid-associated protein that interacts with P, thereby increasing replication accuracy and processivity of the polymerase complex.
Replication of negative-strand RNA viruses relies on two components: a helical ribonucleocapsid and an RNA-dependent RNA polymerase composed of a catalytic subunit, the L protein, and a cofactor, the P protein. We show that the measles virus (MeV) C protein is an additional component of the replication complex. We provide evidence that the C protein is recruited to the ribonucleocapsid by the P protein and map the interacting segments of both C and P proteins. We conclude that the primary function of MeV C is to improve polymerase processivity and accuracy, rather than uniquely to antagonize the type I interferon response. Since most viruses of the
family express C proteins, their primary function may be conserved. |
doi_str_mv | 10.1128/JVI.01733-19 |
format | article |
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, utilizes a complex consisting of genomic RNA, nucleoprotein, the RNA-dependent RNA polymerase, and a polymerase cofactor, the phosphoprotein (P), for transcription and replication. We previously showed that a recombinant MeV that does not express another viral protein, C, has severe transcription and replication deficiencies, including a steeper transcription gradient than the parental virus and generation of defective interfering RNA. This virus is attenuated
and
However, how the C protein operates and whether it is a component of the replication complex remained unclear. Here, we show that C associates with the ribonucleocapsid and forms a complex that can be purified by immunoprecipitation or ultracentrifugation. In the presence of detergent, the C protein is retained on purified ribonucleocapsids less efficiently than the P protein and the polymerase. The C protein is recruited to the ribonucleocapsid through its interaction with the P protein, as shown by immunofluorescence microscopy of cells expressing different combinations of viral proteins and by split luciferase complementation assays. Forty amino-terminal C protein residues are dispensable for the interaction with P, and the carboxyl-terminal half of P is sufficient for the interaction with C. Thus, the C protein, rather than being an "accessory" protein as qualified in textbooks so far, is a ribonucleocapsid-associated protein that interacts with P, thereby increasing replication accuracy and processivity of the polymerase complex.
Replication of negative-strand RNA viruses relies on two components: a helical ribonucleocapsid and an RNA-dependent RNA polymerase composed of a catalytic subunit, the L protein, and a cofactor, the P protein. We show that the measles virus (MeV) C protein is an additional component of the replication complex. We provide evidence that the C protein is recruited to the ribonucleocapsid by the P protein and map the interacting segments of both C and P proteins. We conclude that the primary function of MeV C is to improve polymerase processivity and accuracy, rather than uniquely to antagonize the type I interferon response. Since most viruses of the
family express C proteins, their primary function may be conserved.</description><identifier>ISSN: 0022-538X</identifier><identifier>EISSN: 1098-5514</identifier><identifier>DOI: 10.1128/JVI.01733-19</identifier><identifier>PMID: 31748390</identifier><language>eng</language><publisher>United States: American Society for Microbiology</publisher><subject>Genome and Regulation of Viral Gene Expression ; Life Sciences ; Microbiology and Parasitology ; Spotlight ; Virology</subject><ispartof>Journal of virology, 2020-01, Vol.94 (4)</ispartof><rights>Copyright © 2020 American Society for Microbiology.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><rights>Copyright © 2020 American Society for Microbiology. 2020 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c418t-d131af256ca4ad1dfa40d84e26ec0a3028f392c13668bfdbcaba9a4542c8abf3</citedby><cites>FETCH-LOGICAL-c418t-d131af256ca4ad1dfa40d84e26ec0a3028f392c13668bfdbcaba9a4542c8abf3</cites><orcidid>0000-0001-6881-2548 ; 0000-0001-5539-456X ; 0000-0003-3302-0780 ; 0000-0001-9941-5246</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/PMC6997751/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6997751/$$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/31748390$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-02474276$$DView record in HAL$$Hfree_for_read</backlink></links><search><contributor>Dutch, Rebecca Ellis</contributor><creatorcontrib>Pfaller, Christian K</creatorcontrib><creatorcontrib>Bloyet, Louis-Marie</creatorcontrib><creatorcontrib>Donohue, Ryan C</creatorcontrib><creatorcontrib>Huff, Amanda L</creatorcontrib><creatorcontrib>Bartemes, William P</creatorcontrib><creatorcontrib>Yousaf, Iris</creatorcontrib><creatorcontrib>Urzua, Erica</creatorcontrib><creatorcontrib>Clavière, Mathieu</creatorcontrib><creatorcontrib>Zachary, Marie</creatorcontrib><creatorcontrib>de Masson d'Autume, Valentin</creatorcontrib><creatorcontrib>Carson, Sandra</creatorcontrib><creatorcontrib>Schieferecke, Adam J</creatorcontrib><creatorcontrib>Meyer, Alyssa J</creatorcontrib><creatorcontrib>Gerlier, Denis</creatorcontrib><creatorcontrib>Cattaneo, Roberto</creatorcontrib><title>The C Protein Is Recruited to Measles Virus Ribonucleocapsids by the Phosphoprotein</title><title>Journal of virology</title><addtitle>J Virol</addtitle><description>Measles virus (MeV), like all viruses of the order
, utilizes a complex consisting of genomic RNA, nucleoprotein, the RNA-dependent RNA polymerase, and a polymerase cofactor, the phosphoprotein (P), for transcription and replication. We previously showed that a recombinant MeV that does not express another viral protein, C, has severe transcription and replication deficiencies, including a steeper transcription gradient than the parental virus and generation of defective interfering RNA. This virus is attenuated
and
However, how the C protein operates and whether it is a component of the replication complex remained unclear. Here, we show that C associates with the ribonucleocapsid and forms a complex that can be purified by immunoprecipitation or ultracentrifugation. In the presence of detergent, the C protein is retained on purified ribonucleocapsids less efficiently than the P protein and the polymerase. The C protein is recruited to the ribonucleocapsid through its interaction with the P protein, as shown by immunofluorescence microscopy of cells expressing different combinations of viral proteins and by split luciferase complementation assays. Forty amino-terminal C protein residues are dispensable for the interaction with P, and the carboxyl-terminal half of P is sufficient for the interaction with C. Thus, the C protein, rather than being an "accessory" protein as qualified in textbooks so far, is a ribonucleocapsid-associated protein that interacts with P, thereby increasing replication accuracy and processivity of the polymerase complex.
Replication of negative-strand RNA viruses relies on two components: a helical ribonucleocapsid and an RNA-dependent RNA polymerase composed of a catalytic subunit, the L protein, and a cofactor, the P protein. We show that the measles virus (MeV) C protein is an additional component of the replication complex. We provide evidence that the C protein is recruited to the ribonucleocapsid by the P protein and map the interacting segments of both C and P proteins. We conclude that the primary function of MeV C is to improve polymerase processivity and accuracy, rather than uniquely to antagonize the type I interferon response. Since most viruses of the
family express C proteins, their primary function may be conserved.</description><subject>Genome and Regulation of Viral Gene Expression</subject><subject>Life Sciences</subject><subject>Microbiology and Parasitology</subject><subject>Spotlight</subject><subject>Virology</subject><issn>0022-538X</issn><issn>1098-5514</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNpdkdFP2zAQh61paBTG254nPw6JMJ_tOM7LJFQBLSpaNSq0N8txnMUojYOdVOp_T0oBwZ5OuvvuO51-CH0Dcg5A5c-b-_k5gYyxBPJPaAIkl0maAv-MJoRQmqRM_j1ERzE-EAKcC_4FHTLIuGQ5maC7VW3xFC-D761r8TziP9aEwfW2xL3Ht1bHxkZ878Iwjlzh28E01hvdRVdGXGxxPwqWtY9d7bu95Ss6qHQT7clLPUarq8vVdJYsfl_PpxeLxHCQfVICA13RVBjNdQllpTkpJbdUWEM0I1RWLKcGmBCyqMrC6ELnmqecGqmLih2jX3ttNxRrWxrb9kE3qgturcNWee3Ux0nravXPb5TI8yxLYRSc7gX1f2uzi4Xa9QjlGaeZ2OzYHy_Hgn8cbOzV2kVjm0a31g9RUQYikzQjYkTP9qgJPsZgqzc3ELWLTI2RqefIFOQj_v39G2_wa0bsCct2ktk</recordid><startdate>20200131</startdate><enddate>20200131</enddate><creator>Pfaller, Christian K</creator><creator>Bloyet, Louis-Marie</creator><creator>Donohue, Ryan C</creator><creator>Huff, Amanda L</creator><creator>Bartemes, William P</creator><creator>Yousaf, Iris</creator><creator>Urzua, Erica</creator><creator>Clavière, Mathieu</creator><creator>Zachary, Marie</creator><creator>de Masson d'Autume, Valentin</creator><creator>Carson, Sandra</creator><creator>Schieferecke, Adam J</creator><creator>Meyer, Alyssa J</creator><creator>Gerlier, Denis</creator><creator>Cattaneo, Roberto</creator><general>American Society for Microbiology</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-6881-2548</orcidid><orcidid>https://orcid.org/0000-0001-5539-456X</orcidid><orcidid>https://orcid.org/0000-0003-3302-0780</orcidid><orcidid>https://orcid.org/0000-0001-9941-5246</orcidid></search><sort><creationdate>20200131</creationdate><title>The C Protein Is Recruited to Measles Virus Ribonucleocapsids by the Phosphoprotein</title><author>Pfaller, Christian K ; Bloyet, Louis-Marie ; Donohue, Ryan C ; Huff, Amanda L ; Bartemes, William P ; Yousaf, Iris ; Urzua, Erica ; Clavière, Mathieu ; Zachary, Marie ; de Masson d'Autume, Valentin ; Carson, Sandra ; Schieferecke, Adam J ; Meyer, Alyssa J ; Gerlier, Denis ; Cattaneo, Roberto</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c418t-d131af256ca4ad1dfa40d84e26ec0a3028f392c13668bfdbcaba9a4542c8abf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Genome and Regulation of Viral Gene Expression</topic><topic>Life Sciences</topic><topic>Microbiology and Parasitology</topic><topic>Spotlight</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pfaller, Christian K</creatorcontrib><creatorcontrib>Bloyet, Louis-Marie</creatorcontrib><creatorcontrib>Donohue, Ryan C</creatorcontrib><creatorcontrib>Huff, Amanda L</creatorcontrib><creatorcontrib>Bartemes, William P</creatorcontrib><creatorcontrib>Yousaf, Iris</creatorcontrib><creatorcontrib>Urzua, Erica</creatorcontrib><creatorcontrib>Clavière, Mathieu</creatorcontrib><creatorcontrib>Zachary, Marie</creatorcontrib><creatorcontrib>de Masson d'Autume, Valentin</creatorcontrib><creatorcontrib>Carson, Sandra</creatorcontrib><creatorcontrib>Schieferecke, Adam J</creatorcontrib><creatorcontrib>Meyer, Alyssa J</creatorcontrib><creatorcontrib>Gerlier, Denis</creatorcontrib><creatorcontrib>Cattaneo, Roberto</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</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>Pfaller, Christian K</au><au>Bloyet, Louis-Marie</au><au>Donohue, Ryan C</au><au>Huff, Amanda L</au><au>Bartemes, William P</au><au>Yousaf, Iris</au><au>Urzua, Erica</au><au>Clavière, Mathieu</au><au>Zachary, Marie</au><au>de Masson d'Autume, Valentin</au><au>Carson, Sandra</au><au>Schieferecke, Adam J</au><au>Meyer, Alyssa J</au><au>Gerlier, Denis</au><au>Cattaneo, Roberto</au><au>Dutch, Rebecca Ellis</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The C Protein Is Recruited to Measles Virus Ribonucleocapsids by the Phosphoprotein</atitle><jtitle>Journal of virology</jtitle><addtitle>J Virol</addtitle><date>2020-01-31</date><risdate>2020</risdate><volume>94</volume><issue>4</issue><issn>0022-538X</issn><eissn>1098-5514</eissn><abstract>Measles virus (MeV), like all viruses of the order
, utilizes a complex consisting of genomic RNA, nucleoprotein, the RNA-dependent RNA polymerase, and a polymerase cofactor, the phosphoprotein (P), for transcription and replication. We previously showed that a recombinant MeV that does not express another viral protein, C, has severe transcription and replication deficiencies, including a steeper transcription gradient than the parental virus and generation of defective interfering RNA. This virus is attenuated
and
However, how the C protein operates and whether it is a component of the replication complex remained unclear. Here, we show that C associates with the ribonucleocapsid and forms a complex that can be purified by immunoprecipitation or ultracentrifugation. In the presence of detergent, the C protein is retained on purified ribonucleocapsids less efficiently than the P protein and the polymerase. The C protein is recruited to the ribonucleocapsid through its interaction with the P protein, as shown by immunofluorescence microscopy of cells expressing different combinations of viral proteins and by split luciferase complementation assays. Forty amino-terminal C protein residues are dispensable for the interaction with P, and the carboxyl-terminal half of P is sufficient for the interaction with C. Thus, the C protein, rather than being an "accessory" protein as qualified in textbooks so far, is a ribonucleocapsid-associated protein that interacts with P, thereby increasing replication accuracy and processivity of the polymerase complex.
Replication of negative-strand RNA viruses relies on two components: a helical ribonucleocapsid and an RNA-dependent RNA polymerase composed of a catalytic subunit, the L protein, and a cofactor, the P protein. We show that the measles virus (MeV) C protein is an additional component of the replication complex. We provide evidence that the C protein is recruited to the ribonucleocapsid by the P protein and map the interacting segments of both C and P proteins. We conclude that the primary function of MeV C is to improve polymerase processivity and accuracy, rather than uniquely to antagonize the type I interferon response. Since most viruses of the
family express C proteins, their primary function may be conserved.</abstract><cop>United States</cop><pub>American Society for Microbiology</pub><pmid>31748390</pmid><doi>10.1128/JVI.01733-19</doi><orcidid>https://orcid.org/0000-0001-6881-2548</orcidid><orcidid>https://orcid.org/0000-0001-5539-456X</orcidid><orcidid>https://orcid.org/0000-0003-3302-0780</orcidid><orcidid>https://orcid.org/0000-0001-9941-5246</orcidid><oa>free_for_read</oa></addata></record> |
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title | The C Protein Is Recruited to Measles Virus Ribonucleocapsids by the Phosphoprotein |
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