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{"title"=>"Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm", "type"=>"journal", "authors"=>[{"first_name"=>"Seema S.", "last_name"=>"Lakdawala", "scopus_author_id"=>"55878976100"}, {"first_name"=>"Yicong", "last_name"=>"Wu", "scopus_author_id"=>"8583669600"}, {"first_name"=>"Peter", "last_name"=>"Wawrzusin", "scopus_author_id"=>"55868747400"}, {"first_name"=>"Juraj", "last_name"=>"Kabat", "scopus_author_id"=>"6603934543"}, {"first_name"=>"Andrew J.", "last_name"=>"Broadbent", "scopus_author_id"=>"54779273500"}, {"first_name"=>"Elaine W.", "last_name"=>"Lamirande", "scopus_author_id"=>"6602826730"}, {"first_name"=>"Ervin", "last_name"=>"Fodor", "scopus_author_id"=>"7006539874"}, {"first_name"=>"Nihal", "last_name"=>"Altan-Bonnet", "scopus_author_id"=>"8782813000"}, {"first_name"=>"Hari", "last_name"=>"Shroff", "scopus_author_id"=>"23393654700"}, {"first_name"=>"Kanta", "last_name"=>"Subbarao", "scopus_author_id"=>"7102213212"}], "year"=>2014, "source"=>"PLoS Pathogens", "identifiers"=>{"sgr"=>"84897412935", "doi"=>"10.1371/journal.ppat.1003971", "pui"=>"372738356", "issn"=>"15537374", "pmid"=>"24603687", "isbn"=>"1553-7374 (Electronic)\\r1553-7366 (Linking)", "scopus"=>"2-s2.0-84897412935"}, "id"=>"abaf7e31-85f0-343e-a5bd-c7fe17f13a5d", "abstract"=>"Reassortment of influenza viral RNA (vRNA) segments in co-infected cells can lead to the emergence of viruses with pandemic potential. Replication of influenza vRNA occurs in the nucleus of infected cells, while progeny virions bud from the plasma membrane. However, the intracellular mechanics of vRNA assembly into progeny virions is not well understood. Here we used recent advances in microscopy to explore vRNA assembly and transport during a productive infection. We visualized four distinct vRNA segments within a single cell using fluorescent in situ hybridization (FISH) and observed that foci containing more than one vRNA segment were found at the external nuclear periphery, suggesting that vRNA segments are not exported to the cytoplasm individually. Although many cytoplasmic foci contain multiple vRNA segments, not all vRNA species are present in every focus, indicating that assembly of all eight vRNA segments does not occur prior to export from the nucleus. To extend the observations made in fixed cells, we used a virus that encodes GFP fused to the viral polymerase acidic (PA) protein (WSN PA-GFP) to explore the dynamics of vRNA assembly in live cells during a productive infection. Since WSN PA-GFP colocalizes with viral nucleoprotein and influenza vRNA segments, we used it as a surrogate for visualizing vRNA transport in 3D and at high speed by inverted selective-plane illumination microscopy. We observed cytoplasmic PA-GFP foci colocalizing and traveling together en route to the plasma membrane. Our data strongly support a model in which vRNA segments are exported from the nucleus as complexes that assemble en route to the plasma membrane through dynamic colocalization events in the cytoplasm.", "link"=>"http://www.mendeley.com/research/influenza-virus-assembly-intermediates-fuse-cytoplasm", "reader_count"=>92, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Researcher"=>16, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>40, "Student > Postgraduate"=>2, "Student > Master"=>11, "Student > Bachelor"=>6, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>4}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Researcher"=>16, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>40, "Student > Postgraduate"=>2, "Student > Master"=>11, "Student > Bachelor"=>6, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>14, "Agricultural and Biological Sciences"=>58, "Medicine and Dentistry"=>3, "Veterinary Science and Veterinary Medicine"=>2, "Physics and Astronomy"=>2, "Chemistry"=>2, "Psychology"=>1, "Immunology and Microbiology"=>8}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Chemistry"=>{"Chemistry"=>2}, "Physics and Astronomy"=>{"Physics and Astronomy"=>2}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>8}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>58}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>14}, "Unspecified"=>{"Unspecified"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"United States"=>4, "China"=>1, "Sudan"=>1, "Nigeria"=>1}, "group_count"=>2}

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• {"id"=>"441737102342893569", "text"=>"@PLOSPathogens Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/ZdpMJw0nYf #influenza #coolscienceimages", "created_at"=>"2014-03-07T00:48:26Z", "user"=>"SeemaLakdawala", "user_name"=>"Dr. Seema Lakdawala", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000157748503/dce0496d177649e38b6cb8c22624ea9b_normal.jpeg"}
• {"id"=>"441739531469873152", "text"=>"RT @SeemaLakdawala: @PLOSPathogens Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/ZdpMJw0nYf #influenza #coolsc…", "created_at"=>"2014-03-07T00:58:06Z", "user"=>"TheRealTJWinter", "user_name"=>"TJ Winter", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000628750640/f7cd01b9e192a61a401b295dd69cde73_normal.jpeg"}
• {"id"=>"441963404476305408", "text"=>"@SeemaLakdawala -great job on the PLOS Pathogens paper! http://t.co/0Gx5p2Tqy6", "created_at"=>"2014-03-07T15:47:41Z", "user"=>"DrAndyBroadbent", "user_name"=>"Andrew Broadbent", "user_profile_image"=>"http://pbs.twimg.com/profile_images/3231560527/e10718273059e61df619be15e76d3179_normal.jpeg"}
• {"id"=>"441963697045782528", "text"=>"RT @SeemaLakdawala: @PLOSPathogens Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/ZdpMJw0nYf #influenza #coolsc…", "created_at"=>"2014-03-07T15:48:51Z", "user"=>"DrAndyBroadbent", "user_name"=>"Andrew Broadbent", "user_profile_image"=>"http://pbs.twimg.com/profile_images/3231560527/e10718273059e61df619be15e76d3179_normal.jpeg"}
• {"id"=>"441965779324383232", "text"=>"RT @SeemaLakdawala: Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/slylRrWehZ via @PLOSPathogens #flu", "created_at"=>"2014-03-07T15:57:07Z", "user"=>"IBT_Bioservices", "user_name"=>"IBT Bioservices", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000793809770/82089e9c09707f355ffdf482a90c780f_normal.png"}
• {"id"=>"442012496207962112", "text"=>"#Influenza emergence insight: how virus assembly intermediates fuse in cytoplasm @SeemaLakdawala @NIAIDNews @NIBIBgov http://t.co/j8V5aZNdo9", "created_at"=>"2014-03-07T19:02:45Z", "user"=>"PLOSPathogens", "user_name"=>"PLOS Pathogens", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000667021417/07dfff3f593fc108308ded3df8c96892_normal.jpeg"}
• {"id"=>"442012716463427584", "text"=>"#Influenza reassortment insight: virus assembly intermediates fuse in cytoplasm @SeemaLakdawala @NIAIDNews @NIBIBgov http://t.co/j8V5aZNdo9", "created_at"=>"2014-03-07T19:03:38Z", "user"=>"PLOSPathogens", "user_name"=>"PLOS Pathogens", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000667021417/07dfff3f593fc108308ded3df8c96892_normal.jpeg"}
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• {"id"=>"442034009514975232", "text"=>"RT @PLOSPathogens: #Influenza reassortment insight: virus assembly intermediates fuse in cytoplasm @SeemaLakdawala @NIAIDNews @NIBIBgov htt…", "created_at"=>"2014-03-07T20:28:15Z", "user"=>"SeemaLakdawala", "user_name"=>"Dr. Seema Lakdawala", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000157748503/dce0496d177649e38b6cb8c22624ea9b_normal.jpeg"}
• {"id"=>"442048444845146112", "text"=>"#PLOSPathogens: Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/D3RtG90bya", "created_at"=>"2014-03-07T21:25:36Z", "user"=>"shadigeravandi", "user_name"=>"Shadi Geravandi", "user_profile_image"=>"http://pbs.twimg.com/profile_images/439213812865323008/1yphME2X_normal.jpeg"}
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• {"id"=>"446313370895921152", "text"=>"3D-localization of four distinct vRNA segments using iSPIM and Imaris http://t.co/917edSAaPh http://t.co/C9d5mKJxVv", "created_at"=>"2014-03-19T15:52:54Z", "user"=>"lucianoaglucas", "user_name"=>"Luciano Lucas", "user_profile_image"=>"http://pbs.twimg.com/profile_images/444854482891837440/5ypLd3R8_normal.jpeg"}
• {"id"=>"446314395975446528", "text"=>"3D-localization of four distinct vRNA segments using iSPIM and Imaris http://t.co/917edSAaPh #microscopy #spim #3D http://t.co/4YNu67w1Iw", "created_at"=>"2014-03-19T15:56:59Z", "user"=>"lucianoaglucas", "user_name"=>"Luciano Lucas", "user_profile_image"=>"http://pbs.twimg.com/profile_images/444854482891837440/5ypLd3R8_normal.jpeg"}
• {"id"=>"446717266772496384", "text"=>"Cytoplasm provides a workshop for influenza #virus RNA to reassort & assemble:\nhttp://t.co/C7evwve5Nq @PLOSPathogens http://t.co/2R7duj918L", "created_at"=>"2014-03-20T18:37:50Z", "user"=>"StojdlLab", "user_name"=>"Stojdl Lab", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000727825532/bc4a9962f88946130c8ffaa016b00604_normal.jpeg"}
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• {"id"=>"446801892387991552", "text"=>"Influenza A virus assembly intermediates fuse in the cytoplasm http://t.co/JAUGWOip9c - beautiful imaging @drandybroadbent tell Kanta v nice", "created_at"=>"2014-03-21T00:14:06Z", "user"=>"10queues", "user_name"=>"Paul Duprex", "user_profile_image"=>"http://pbs.twimg.com/profile_images/2078166162/paul_normal.jpg"}
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• {"id"=>"448569946994262016", "text"=>"RT @StojdlLab: Cytoplasm provides a workshop for influenza #virus RNA to reassort & assemble:\nhttp://t.co/C7evwve5Nq @PLOSPathogens http://…", "created_at"=>"2014-03-25T21:19:44Z", "user"=>"busters13okd", "user_name"=>"o-kazuma", "user_profile_image"=>"http://pbs.twimg.com/profile_images/2986538744/b207a5120ecec603c7c8a8458c5eb14f_normal.jpeg"}
• {"id"=>"449312263342522368", "text"=>"March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA http://t.co/VG99uijoYq", "created_at"=>"2014-03-27T22:29:26Z", "user"=>"PLOSPathogens", "user_name"=>"PLOS Pathogens", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000667021417/07dfff3f593fc108308ded3df8c96892_normal.jpeg"}
• {"id"=>"449312765208199168", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T22:31:25Z", "user"=>"MicrobLog_me_uk", "user_name"=>"MicrobLog™", "user_profile_image"=>"http://pbs.twimg.com/profile_images/352341447/microblogo2008_normal.jpg"}
• {"id"=>"449313358047502336", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T22:33:47Z", "user"=>"LordBexar", "user_name"=>"Paul DeOrd", "user_profile_image"=>"http://pbs.twimg.com/profile_images/274618005/Me_on_Velour_normal.jpg"}
• {"id"=>"449315350593957888", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T22:41:42Z", "user"=>"ViralPseudotype", "user_name"=>"ViralPseudotypeUnit", "user_profile_image"=>"http://pbs.twimg.com/profile_images/1891375569/3D_Influenza_blue_no_key_full_sml2_normal.jpg"}
• {"id"=>"449317171613925376", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T22:48:56Z", "user"=>"DrSiYoung", "user_name"=>"Simon Young", "user_profile_image"=>"http://pbs.twimg.com/profile_images/412350032638005248/ZkHySvsD_normal.jpeg"}
• {"id"=>"449325497835474944", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T23:22:01Z", "user"=>"PLOS", "user_name"=>"PLOS", "user_profile_image"=>"http://pbs.twimg.com/profile_images/2424363764/2etoq0zjwxicokm1woge_normal.jpeg"}
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• {"id"=>"449325659350118401", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T23:22:39Z", "user"=>"sev_virologia", "user_name"=>"SEV Virología", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000722169773/9967afd82f734acf4066ad49324d0193_normal.jpeg"}
• {"id"=>"449325808201773056", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T23:23:15Z", "user"=>"TheDoggieDaddy", "user_name"=>"Doggie Daddy", "user_profile_image"=>"http://pbs.twimg.com/profile_images/446818481191649281/-Ac9F5nX_normal.jpeg"}
• {"id"=>"449331229822054400", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-27T23:44:48Z", "user"=>"SarahRLeist", "user_name"=>"Sarah Я Leist", "user_profile_image"=>"http://pbs.twimg.com/profile_images/412302985637527552/cV9PEQEN_normal.jpeg"}
• {"id"=>"449337004958838785", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T00:07:44Z", "user"=>"shyduroff", "user_name"=>"shyduroff", "user_profile_image"=>"http://pbs.twimg.com/profile_images/3695201339/dcec331712656e45a8bbeebc392e5ec8_normal.jpeg"}
• {"id"=>"449337652614266881", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T00:10:19Z", "user"=>"johnmgonzalez", "user_name"=>"John M. Gonzalez", "user_profile_image"=>"http://pbs.twimg.com/profile_images/2941792605/bb9bf211971029386b3f0b70776982a1_normal.jpeg"}
• {"id"=>"449338471862525952", "text"=>"Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm. http://t.co/sbfWiDs00v", "created_at"=>"2014-03-28T00:13:34Z", "user"=>"BIOCIENCIA2013", "user_name"=>"BIOCIENCIA", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000300217090/9b25d5f581a95fff46afd4d8d0d0aad0_normal.jpeg"}
• {"id"=>"449339355384254464", "text"=>"RT @BIOCIENCIA2013: Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm. http://t.co/sbfWiDs00v", "created_at"=>"2014-03-28T00:17:05Z", "user"=>"Vakdaro", "user_name"=>"Daniel Romero", "user_profile_image"=>"http://pbs.twimg.com/profile_images/443089177538617344/W-sWD8Jn_normal.jpeg"}
• {"id"=>"449350784170594306", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T01:02:30Z", "user"=>"SeemaLakdawala", "user_name"=>"Dr. Seema Lakdawala", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000157748503/dce0496d177649e38b6cb8c22624ea9b_normal.jpeg"}
• {"id"=>"449353443644571648", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T01:13:04Z", "user"=>"MyNameIsJanineB", "user_name"=>"Janine B", "user_profile_image"=>"http://pbs.twimg.com/profile_images/449342899658842112/xDZWjtIy_normal.jpeg"}
• {"id"=>"449356499832561664", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T01:25:12Z", "user"=>"TheRealTJWinter", "user_name"=>"TJ Winter", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000628750640/f7cd01b9e192a61a401b295dd69cde73_normal.jpeg"}
• {"id"=>"449526952647720960", "text"=>"Very nice work by @SeemaLakdawala @PLOSPathogens shedding light on mechanism behind influenza viral RNA reassortment http://t.co/CiHgfiQUFa", "created_at"=>"2014-03-28T12:42:31Z", "user"=>"bansallab", "user_name"=>"Bansal Lab", "user_profile_image"=>"http://pbs.twimg.com/profile_images/2505322529/b5fgspytr6m299uxjbd9_normal.png"}
• {"id"=>"449527103080644608", "text"=>"RT @bansallab: Very nice work by @SeemaLakdawala @PLOSPathogens shedding light on mechanism behind influenza viral RNA reassortment http://…", "created_at"=>"2014-03-28T12:43:07Z", "user"=>"BioAndBaseball", "user_name"=>"Matt Watson", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000447121930/1cd0e3c6ce6711fe404b3c93a9325a55_normal.jpeg"}
• {"id"=>"449539264477679616", "text"=>"RT @bansallab: Very nice work by @SeemaLakdawala @PLOSPathogens shedding light on mechanism behind influenza viral RNA reassortment http://…", "created_at"=>"2014-03-28T13:31:27Z", "user"=>"SeemaLakdawala", "user_name"=>"Dr. Seema Lakdawala", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000157748503/dce0496d177649e38b6cb8c22624ea9b_normal.jpeg"}
• {"id"=>"449608592187219969", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T18:06:56Z", "user"=>"DrAndyBroadbent", "user_name"=>"Andrew Broadbent", "user_profile_image"=>"http://pbs.twimg.com/profile_images/3231560527/e10718273059e61df619be15e76d3179_normal.jpeg"}
• {"id"=>"449609023281954816", "text"=>"RT @PLOSPathogens: March cover: a 3D rendering of an MDCK cell infected with H1N1 @NIAIDnews @SeemaLakdawala @NIH http://t.co/xqHhX6U2bA ht…", "created_at"=>"2014-03-28T18:08:39Z", "user"=>"EstherBlanco01", "user_name"=>"Esther Blanco", "user_profile_image"=>"http://pbs.twimg.com/profile_images/2412676485/jlrzlay0eggkqbjv8xwn_normal.jpeg"}
• {"id"=>"451162259247874048", "text"=>"RT @lucianoaglucas: 3D-localization of four distinct vRNA segments using iSPIM and Imaris http://t.co/917edSAaPh #microscopy #spim #3D http…", "created_at"=>"2014-04-02T01:00:39Z", "user"=>"SeemaLakdawala", "user_name"=>"Dr. Seema Lakdawala", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000157748503/dce0496d177649e38b6cb8c22624ea9b_normal.jpeg"}
• {"id"=>"452385806590541824", "text"=>"PLOS Pathogens: Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/wHMTtsgkRQ (Sourced by http://t.co/0rVypuOZ0C)", "created_at"=>"2014-04-05T10:02:35Z", "user"=>"H7N9Influenza", "user_name"=>"Dave Roberts", "user_profile_image"=>"http://pbs.twimg.com/profile_images/3501363698/67e86cb1c0a5dbe79300ad64694b8c9d_normal.jpeg"}
• {"id"=>"452559046533738496", "text"=>"RT @H7N9Influenza: PLOS Pathogens: Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/wHMTtsgkRQ (Sourced by http:/…", "created_at"=>"2014-04-05T21:30:59Z", "user"=>"BaronianConsult", "user_name"=>"Baronian Consulting", "user_profile_image"=>"http://pbs.twimg.com/profile_images/344513261580112136/1bef8d5247ba652c1f6a57a1311172d0_normal.jpeg"}
• {"id"=>"453227092079951872", "text"=>"RT @H7N9Influenza: PLOS Pathogens: Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm http://t.co/wHMTtsgkRQ (Sourced by http:/…", "created_at"=>"2014-04-07T17:45:33Z", "user"=>"PLOSPathogens", "user_name"=>"PLOS Pathogens", "user_profile_image"=>"http://pbs.twimg.com/profile_images/378800000667021417/07dfff3f593fc108308ded3df8c96892_normal.jpeg"}
• {"id"=>"644909214079561728", "text"=>"From @SeemaLakdawala recent article (http://t.co/kL1Ss7xYr7) shows flu genome segments form intermediates that fuse en route.", "created_at"=>"2015-09-18T16:21:53Z", "user"=>"vscooper", "user_name"=>"Vaughn Cooper", "user_profile_image"=>"http://pbs.twimg.com/profile_images/603744672868077568/aTZJfiuH_normal.jpg"}

Wikipedia | Further Information

• {"title"=>"File:Influenza-A-Virus-Assembly-Intermediates-Fuse-in-the-Cytoplasm-ppat.1003971.s009.ogv", "url"=>"http://commons.wikimedia.org/wiki/File:Influenza-A-Virus-Assembly-Intermediates-Fuse-in-the-Cytoplasm-ppat.1003971.s009.ogv", "timestamp"=>"2018-01-13T08:27:40Z"}
• {"title"=>"File:Influenza-A-Virus-Assembly-Intermediates-Fuse-in-the-Cytoplasm-ppat.1003971.s008.ogv", "url"=>"http://commons.wikimedia.org/wiki/File:Influenza-A-Virus-Assembly-Intermediates-Fuse-in-the-Cytoplasm-ppat.1003971.s008.ogv", "timestamp"=>"2018-01-13T08:27:39Z"}
• {"title"=>"File:Influenza-A-Virus-Assembly-Intermediates-Fuse-in-the-Cytoplasm-ppat.1003971.s007.ogv", "url"=>"http://commons.wikimedia.org/wiki/File:Influenza-A-Virus-Assembly-Intermediates-Fuse-in-the-Cytoplasm-ppat.1003971.s007.ogv", "timestamp"=>"2018-01-13T08:27:38Z"}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/1410690", "https://ndownloader.figshare.com/files/1410691", "https://ndownloader.figshare.com/files/1410692", "https://ndownloader.figshare.com/files/1410693", "https://ndownloader.figshare.com/files/1410694", "https://ndownloader.figshare.com/files/1410695", "https://ndownloader.figshare.com/files/1410696", "https://ndownloader.figshare.com/files/1410697", "https://ndownloader.figshare.com/files/1410698", "https://ndownloader.figshare.com/files/1410699", "https://ndownloader.figshare.com/files/1410700", "https://ndownloader.figshare.com/files/1410701"], "description"=>"<div><p>Reassortment of influenza viral RNA (vRNA) segments in co-infected cells can lead to the emergence of viruses with pandemic potential. Replication of influenza vRNA occurs in the nucleus of infected cells, while progeny virions bud from the plasma membrane. However, the intracellular mechanics of vRNA assembly into progeny virions is not well understood. Here we used recent advances in microscopy to explore vRNA assembly and transport during a productive infection. We visualized four distinct vRNA segments within a single cell using fluorescent <i>in situ</i> hybridization (FISH) and observed that foci containing more than one vRNA segment were found at the external nuclear periphery, suggesting that vRNA segments are not exported to the cytoplasm individually. Although many cytoplasmic foci contain multiple vRNA segments, not all vRNA species are present in every focus, indicating that assembly of all eight vRNA segments does not occur prior to export from the nucleus. To extend the observations made in fixed cells, we used a virus that encodes GFP fused to the viral polymerase acidic (PA) protein (WSN PA-GFP) to explore the dynamics of vRNA assembly in live cells during a productive infection. Since WSN PA-GFP colocalizes with viral nucleoprotein and influenza vRNA segments, we used it as a surrogate for visualizing vRNA transport in 3D and at high speed by inverted selective-plane illumination microscopy. We observed cytoplasmic PA-GFP foci colocalizing and traveling together en route to the plasma membrane. Our data strongly support a model in which vRNA segments are exported from the nucleus as complexes that assemble en route to the plasma membrane through dynamic colocalization events in the cytoplasm.</p></div>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "intermediates", "fuse"], "article_id"=>954398, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1003971.s001", "https://dx.doi.org/10.1371/journal.ppat.1003971.s002", "https://dx.doi.org/10.1371/journal.ppat.1003971.s003", "https://dx.doi.org/10.1371/journal.ppat.1003971.s004", "https://dx.doi.org/10.1371/journal.ppat.1003971.s005", "https://dx.doi.org/10.1371/journal.ppat.1003971.s006", "https://dx.doi.org/10.1371/journal.ppat.1003971.s007", "https://dx.doi.org/10.1371/journal.ppat.1003971.s008", "https://dx.doi.org/10.1371/journal.ppat.1003971.s009", "https://dx.doi.org/10.1371/journal.ppat.1003971.s010", "https://dx.doi.org/10.1371/journal.ppat.1003971.s011", "https://dx.doi.org/10.1371/journal.ppat.1003971.s012"], "stats"=>{"downloads"=>13, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Influenza_A_Virus_Assembly_Intermediates_Fuse_in_the_Cytoplasm_/954398", "title"=>"Influenza A Virus Assembly Intermediates Fuse in the Cytoplasm", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410650"], "description"=>"<p>Fluorescent <i>in situ</i> hybridization (FISH) for viral RNA segments 1 (PB2) and 4 (HA) on MDCK (A) or A549 cells (B) cells 16 hpi with WSN PA-GFP (MOI = 0.5). Images to the right are enlarged regions identified by the dashed box. White arrowheads show colocalization of PA-GFP and vRNA, and open arrowheads indicate GFP foci not colocalized with viral RNA signal. DAPI marks the cellular nucleus. Scale bars are 5 µm in all panels of part A. In part B scale bars are 10 µm in the whole cell images and 2 µm in length in the enlarged region.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "wsn", "pa-gfp", "vrna"], "article_id"=>954364, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g005", "stats"=>{"downloads"=>3, "page_views"=>82, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Colocalization_of_WSN_PA_GFP_and_influenza_vRNA_segments_/954364", "title"=>"Colocalization of WSN PA-GFP and influenza vRNA segments.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410640"], "description"=>"<p>Schematic of the reverse genetics pHW2000 plasmid created by inserting the coding region of full length GFP into the C-terminal domain of the WSN PA open reading frame (A). This insertion included a duplication of 162 nucleotides of WSN PA prior to the 5′ non-coding region (NCR) depicted in the black rectangle. Western blot of purified recombinant WT WSN or WSN PA-GFP virus grown in embryonated eggs (B). The GFP signal corresponds to the appropriate size of a PA-GFP fusion protein. Viral NP protein was used as a control to ensure loading of equivalent amounts of virus. Immunofluorescence of MDCK cells infected with WSN PA-GFP (MOI = 3) for 16 hpi with anti-influenza NP antibody (C). Images to the right are enlarged regions identified by the dashed square. White arrows show areas of co-localization. All scale bars are 5 µm.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "wsn", "pa-gfp"], "article_id"=>954361, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g004", "stats"=>{"downloads"=>4, "page_views"=>37, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Generation_of_WSN_PA_GFP_influenza_virus_/954361", "title"=>"Generation of WSN PA-GFP influenza virus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410638"], "description"=>"<p>3D rendering of an MDCK cell infected with WT WSN (MOI = 3) for 8 hpi were stained with probe B from <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003971#ppat.1003971.s010\" target=\"_blank\">Table S1</a>: WSN PB2 Quasar 570 (red spots), PB1 Alexa-Fluor 488 (green spots), PA Cal Fluor Red 590 (orange spots), and NP Quasar 670 (yellow spots) (A). The DAPI-stained nucleus is labeled in blue. An enlarged region of the 3D image was tilted 90° in the z-direction to provide an axial-view of the cytoplasmic foci and nucleus. Scale bars are 4 µm in the whole cell image and 2 µm in the rotated images. The total number of foci containing 1, 2, 3, or 4 vRNA segments was quantified within a given cell (B). Each bar represents the average from 3 independently analyzed cells with standard error indicated. The distance from the nucleus for each focus from four independent cells was calculated (C). The proportion of foci containing 4,3,2,or 1 distinct vRNA segment with a given range from the nucleus is represented graphically as a scatter plot. Each spot is an average from 4 independent cells and the standard error is indicated.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "rna", "spatial", "cytoplasmic"], "article_id"=>954359, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g002", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Viral_RNA_composition_and_spatial_location_of_cytoplasmic_foci_/954359", "title"=>"Viral RNA composition and spatial location of cytoplasmic foci.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410639"], "description"=>"<p>Graphical representation of the Pearson correlation coefficient values between each vRNA segment and all other vRNA segments from exclusively cytoplasmic staining. Dual staining of infected cells with FISH probes against the same target served as a positive control and is indicated in red. Each spot represents a single analyzed infected cell. Gray shading highlights the region of low colocalization based on the Pearson correlation coefficient.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "coefficients", "vrna"], "article_id"=>954360, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g003", "stats"=>{"downloads"=>4, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Colocalization_coefficients_between_influenza_vRNA_segments_/954360", "title"=>"Colocalization coefficients between influenza vRNA segments.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410637"], "description"=>"<p>MDCK cells were infected with wild type (WT) WSN at a multiplicity of infection (MOI) of 3 for 8 hours post infection (hpi) and then imaged with FISH probes directed against four distinct vRNA species. The first row displays a cell probed for influenza vRNA segments PB2, PB1, PA, and NP. Row 3 contains images of a cell probed for the other 4 influenza vRNA segments: HA, NA, M and NS. The second and fourth row of images show an enlargement of the area defined by the dashed boxes in the first and third rows. Solid white arrowheads identify a cytoplasmic focus with all four distinct vRNA segments, turquoise arrowheads indicate a focus with only three vRNA species, and open arrowheads show a focus with only two vRNA species. All scale bars are 10 µm. DAPI marks the cellular nucleus.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "viral", "segments"], "article_id"=>954358, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g001", "stats"=>{"downloads"=>2, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Visualization_of_four_distinct_influenza_viral_gene_segments_within_a_single_cell_/954358", "title"=>"Visualization of four distinct influenza viral gene segments within a single cell.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410655"], "description"=>"<p>The data presented in this paper supports a model of vRNA assembly whereby 1) vRNA segments export from the nucleus as complexes containing multiple vRNA segments, 2) cytoplasmic foci containing vRNA segments transport through the cytoplasm and 3) these cytoplasmic foci fuse together in the cytoplasm prior to budding. Each vRNA segment is shown as a loop where the ends are associated with a heterotrimeric polymerase complex identified as 3 circles. The PA protein of the polymerase complex is shown in green since this was the protein used as a surrogate for vRNA transport. The cellular Rab11a-containing endosomes that colocalize with vRNA-containing cytoplasmic foci are indicated.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "vrna"], "article_id"=>954369, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g007", "stats"=>{"downloads"=>5, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proposed_model_for_vRNA_assembly_/954369", "title"=>"Proposed model for vRNA assembly.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}
• {"files"=>["https://ndownloader.figshare.com/files/1410653"], "description"=>"<p>Maximum intensity projection from inverted selective-plane illumination microscopy (iSPIM) movie of MDCK cells infected with WSN PA-GFP (MOI = 5) at 16 hpi, 36 seconds (s) after onset of tracking (A). The red-boxed region is highlighted in (B and C), and the red star indicates the region where colocalization of two GFP foci (marked in red and blue arrows) occurred. A single WSN PA-GFP focus trajectory; each red dot is a spot tracked in time, the yellow line is the overall trace, and white arrows indicate the direction of the PA-GFP movement (B). The total track duration is 46 s. Part C contains still frames of the red-boxed region in part A from 26–44 s, and illustrates colocalization and subsequent fusion of two foci. The red arrow indicates the PA-GFP focus that is being tracked and the blue arrow indicates the PA-GFP focus with which it fuses. All scale bars are 2 µm. An intensity versus depth plot of the colocalized foci, indicated by the red star in part A, demonstrates a single peak indicating that the fusion occurs in a single diffraction-limited spot (D). An intensity vs. time plot of the fused focus compared to an adjacent stationary focus, identified by the white circle in part C, during the same time frame (E). The asterisk marks the time corresponding to the colocalization event.</p>", "links"=>[], "tags"=>["Infectious diseases", "Viral diseases", "influenza", "wsn", "pa-gfp", "cytoplasm", "mdck"], "article_id"=>954367, "categories"=>["Medicine"], "users"=>["Seema S. Lakdawala", "Yicong Wu", "Peter Wawrzusin", "Juraj Kabat", "Andrew J. Broadbent", "Elaine W. Lamirande", "Ervin Fodor", "Nihal Altan-Bonnet", "Hari Shroff", "Kanta Subbarao"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003971.g006", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fusion_event_of_an_WSN_PA_GFP_focus_in_the_cytoplasm_of_MDCK_cells_/954367", "title"=>"Fusion event of an WSN PA-GFP focus in the cytoplasm of MDCK cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 04:10:56"}

PMC Usage Stats | Further Information

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Relative Metric

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F1000Prime