De Novo Synthesis of VP16 Coordinates the Exit from HSV Latency In Vivo
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{"title"=>"De novo synthesis of VP16 coordinates the exit from HSV latency in vivo", "type"=>"journal", "authors"=>[{"first_name"=>"Richard L.", "last_name"=>"Thompson", "scopus_author_id"=>"55482219200"}, {"first_name"=>"Chris M.", "last_name"=>"Preston", "scopus_author_id"=>"7202709397"}, {"first_name"=>"Nancy M.", "last_name"=>"Sawtell", "scopus_author_id"=>"7004060497"}], "year"=>2009, "source"=>"PLoS Pathogens", "identifiers"=>{"pmid"=>"19325890", "doi"=>"10.1371/journal.ppat.1000352", "sgr"=>"63449116249", "isbn"=>"1553-7374 (Electronic)\\n1553-7366 (Linking)", "scopus"=>"2-s2.0-63449116249", "issn"=>"15537366", "pui"=>"354404896"}, "id"=>"963d60a9-6103-3445-8b78-de927426a719", "abstract"=>"The mechanism controlling the exit from herpes simplex virus latency (HSV) is of central importance to recurrent disease and transmission of infection, yet interactions between host and viral functions that govern this process remain unclear. The cascade of HSV gene transcription is initiated by the multifunctional virion protein VP16, which is expressed late in the viral replication cycle. Currently, it is widely accepted that VP16 transactivating function is not involved in the exit from latency. Utilizing the mouse ocular model of HSV pathogenesis together with genetically engineered viral mutants and assays to quantify latency and the exit from latency at the single neuron level, we show that in vivo (i) the VP16 promoter confers distinct regulation critical for viral replication in the trigeminal ganglion (TG) during the acute phase of infection and (ii) the transactivation function of VP16 (VP16TF) is uniquely required for the exit from latency. TG neurons latently infected with the VP16TF mutant in1814 do not express detectable viral proteins following stress, whereas viruses with mutations in the other major viral transcription regulators ICP0 and ICP4 do exit the latent state. Analysis of a VP16 promoter/reporter mutant in the background of in1814 demonstrates that the VP16 promoter is activated in latently infected neurons following stress in the absence of other viral proteins. These findings support the novel hypothesis that de novo expression of VP16 regulates entry into the lytic program in neurons at all phases of the viral life cycle. HSV reactivation from latency conforms to a model in which stochastic derepression of the VP16 promoter and expression of VP16 initiates entry into the lytic cycle.", "link"=>"http://www.mendeley.com/research/novo-synthesis-vp16-coordinates-exit-hsv-latency-vivo", "reader_count"=>51, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>9, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>7, "Student > Bachelor"=>7, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>9, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>14, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>7, "Student > Bachelor"=>7, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>33, "Medicine and Dentistry"=>8, "Neuroscience"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Physics and Astronomy"=>1, "Chemistry"=>1, "Immunology and Microbiology"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>8}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>33}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Colombia"=>1, "Belgium"=>1, "United States"=>2, "Chile"=>1, "Germany"=>1}, "group_count"=>1}

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  • {"files"=>["https://ndownloader.figshare.com/files/903634"], "description"=>"<p>(A) Mice were infected with 17VP16pLZ. At the indicated days pi, TG were\n removed and sequentially processed for the detection of b-gal activity\n (blue) and viral proteins (purple), as detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. A photomicrograph of a focus of infection\n in a TG is shown. White arrows indicate neurons with b-gal activity with\n little or no detectable viral protein. Counts of neurons positive for\n only b-gal or b-gal plus viral protein are indicated below the\n micrograph. (B) Photomicrographs of viral plaques on RSC monolayers\n infected at low moi with 17-0pZ563gJ and 17VP16pLZ are shown. The\n monolayers were stained to detect b-gal and viral proteins. At regions\n at the edge of the plaques formed by 17-0pZ563gJ cells expressing b-gal\n (blue) with little or no evidence of viral protein expression were\n evident (arrow). Regions at the edge of plaques formed by 17VP16pLZ show\n cells staining positive for viral proteins (purple) with little or no\n staining for b-gal evident (arrow).</p>", "links"=>[], "tags"=>["vp16", "promoter", "sensory", "neurons", "acute"], "article_id"=>574092, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Activation_of_the_VP16_promoter_in_sensory_neurons_during_acute____infection_/574092", "title"=>"Activation of the VP16 promoter in sensory neurons during acute\n infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:27:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/904186"], "description"=>"<p>(A) Shown is a schematic of the role of de novo VP16 expression during acute\n infection of the trigeminal ganglion. As detailed in the text, we\n hypothesize that VP16 protein is not transported efficiently to neuronal\n cell bodies and that sequences present in the VP16 promoter direct the de\n novo expression of VP16, which overcomes repressors (e.g. riboregulators) to\n initiate lytic infection during the acute stage of viral replication. (B) We\n hypothesize that following stress the de novo synthesis of VP16 initiates a\n feedback loop with the IE genes that results in viral reactivation in one or\n a very few of the 6,000 latently infected neurons present in the\n ganglion.</p>", "links"=>[], "tags"=>["vp16", "controls", "lytic", "latent", "sensory", "neurons", "stages", "infection"], "article_id"=>574633, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g007", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_regulation_of_expression_of_VP16_controls_the_balance_between_lytic____and_latent_infection_of_sensory_neurons_at_all_stages_of_the_virus_infection____cycle_/574633", "title"=>"The regulation of expression of VP16 controls the balance between lytic\n and latent infection of sensory neurons at all stages of the virus infection\n cycle.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:29:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/904061"], "description"=>"<p>(A) Mice latently infected with the indicated strains were subjected to\n HS, and whole TG were assayed for viral protein positive cells 22 hrs\n post stress. Each point in the scattergram represents the number of\n viral protein positive neurons detected in an individual TG. The\n horizontal bars are drawn at the mean. (B) A photomicrograph of a whole\n mounted ganglion processed for the detection of HSV-1 proteins 22 hr\n post HS shows a single neuron in which the latent genome has entered the\n lytic cycle. The brown precipitate evident in the cell body and axonal\n tract indicates the presence of viral proteins.</p>", "links"=>[], "tags"=>["latency"], "article_id"=>574522, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g006", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In1814_does_not_exit_latency_in_vivo_/574522", "title"=>"In1814 does not exit latency in vivo.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:29:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/903966"], "description"=>"<p>(A) Mice were infected as described in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. At 40 days pi, latently infected mice were induced\n to reactivate using HS, and TG were removed and assayed for infectious\n virus as detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. The\n histograms represent the percentage of mice in which reactivated HSV was\n detected. The number of mice positive/tested is shown on the histograms.\n (B) Explant reactivation of ganglia latently infected with in1814 or\n 17VP16Δ422. TG from latently infected mice were axotomized and\n explanted. In1814 and 17VP16Δ422 infected TG were assayed for\n infectious virus on days 5 and 15 post explant, respectively. The GFP\n expression cassette in 17VP16Δ422 was used to detect gene\n expression from the viral genome during explantation. Exit from latency\n was observed as single GFP positive neuron at day 4 (C) which resulted\n in the spread of virus through the TG by day 6 (D).</p>", "links"=>[], "tags"=>["mutant", "in1814", "reactivate", "latency", "vivo", "does", "explanted"], "article_id"=>574421, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_mutant_in1814_does_not_reactivate_from_latency_in_vivo_but_does____reactivate_in_explanted_ganglia_/574421", "title"=>"The mutant in1814 does not reactivate from latency in vivo but does\n reactivate in explanted ganglia.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:28:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/903447"], "description"=>"<p>(A) A diagram of the construction of the VP5p/VP16 mutants is shown. The\n VP16 promoter and 5′UTR sequences were replaced with those of\n another gene expressed with leaky late kinetics, the VP5 gene, as\n detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. (B) RSC were\n infected with mutants VP5/VP16-1 and -3, the genomically restored\n isolate (VP5p/VP16-1R), and wild type HSV-1 strain 17Syn+ at an\n moi of 0.0004 pfu/cell. At the indicated times, 3 plates infected with\n each virus were harvested and assayed independently for virus content as\n detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. (C,D) Mice\n were infected as detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a> and, at the indicated times pi, tissues from three mice\n from each group were assayed for virus content. The grey shading in C\n and D indicates the regions employed to calculate the areas under the\n curves for the VP5p/VP16 mutants.</p>", "links"=>[], "tags"=>["mutants", "vitro"], "article_id"=>573903, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g001", "stats"=>{"downloads"=>3, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Replication_of_VP5p_VP16_mutants_in_vitro_and_in_vivo_/573903", "title"=>"Replication of VP5p/VP16 mutants in vitro and in vivo.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:26:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/903853"], "description"=>"<p>Groups of mice were infected with strain 17Syn+, in1814, and the\n genomically rescued variant 1814R, as described in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. At 40 days pi, the ganglia of 3 mice per\n group were processed for single neuron PCR. Individual neurons were\n examined for the presence of the viral genome and the number of viral\n genomes present in positive neurons was determined using a quantitative\n PCR assay as detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>.\n (A) Shown is the percentage of neurons positive for the viral genome.\n The number of neurons positive for the viral genome over the number\n tested is shown in the histograms. (B) Each point on the scattergram\n represents the number of viral genomes present in an individual neuron.\n The horizontal bars are drawn at the mean value of genome copies per\n positive neuron.</p>", "links"=>[], "tags"=>["in1814", "establishes", "latency", "efficiently", "genomically", "wild"], "article_id"=>574309, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g004", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutant_in1814_establishes_latency_as_efficiently_as_genomically_wild____type_isolates_/574309", "title"=>"Mutant in1814 establishes latency as efficiently as genomically wild\n type isolates.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:28:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/903756"], "description"=>"<p>(A) A schematic of the experimental design employed to biologically\n characterize the VP16 mutants is shown. (B) A diagram of the genomic\n structures of in1814 and 17VP16Δ422 is shown. (C,D) Mice were\n infected as detailed in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>\n and, at the indicated times pi, tissues from three mice from each group\n were assayed for infectious virus. Solid lines represent titers obtained\n by standard plaque assay, whereas dashed lines represent the same\n samples titrated in the presence of 3 mM HMBA, as described in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000352#s4\" target=\"_blank\">Methods</a>. The area under the curve\n (AUC) was calculated for each curve, and the fold increase in AUC in the\n presence of HMBA is indicated below the graphs. Genomically restored\n isolates of both in1814 (1814R) and 17VP16Δ422\n (17VP16Δ422R) were analyzed and found to be not different from\n the parental strain, 17syn+ (not shown).</p>", "links"=>[], "tags"=>["kinetics", "vp16", "transactivation", "mutants", "in1814", "and"], "article_id"=>574209, "categories"=>["Medicine", "Infectious Diseases", "Virology", "Neuroscience"], "users"=>["Richard L. Thompson", "Chris M. Preston", "Nancy M. Sawtell"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1000352.g003", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Replication_kinetics_of_the_VP16_transactivation_mutants_in1814_and____17VP16_916_422_/574209", "title"=>"Replication kinetics of the VP16 transactivation mutants in1814 and\n 17VP16Δ422.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 06:27:43"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"9", "full-text"=>"10", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}

Relative Metric

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