Selective Histonedeacetylase Inhibitor M344 Intervenes in HIV-1 Latency through Increasing Histone Acetylation and Activation of NF-kappaB
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{"title"=>"Selective Histonedeacetylase Inhibitor M344 Intervenes in HIV-1 Latency through Increasing Histone Acetylation and Activation of NF-kappaB", "type"=>"journal", "authors"=>[{"first_name"=>"Hao", "last_name"=>"Ying", "scopus_author_id"=>"36605392300"}, {"first_name"=>"Yuhao", "last_name"=>"Zhang", "scopus_author_id"=>"56721059300"}, {"first_name"=>"Xin", "last_name"=>"Zhou", "scopus_author_id"=>"56228304100"}, {"first_name"=>"Xiying", "last_name"=>"Qu", "scopus_author_id"=>"55487309600"}, {"first_name"=>"Pengfei", "last_name"=>"Wang", "scopus_author_id"=>"7405458795"}, {"first_name"=>"Sijie", "last_name"=>"Liu", "scopus_author_id"=>"37097599300"}, {"first_name"=>"Daru", "last_name"=>"Lu", "scopus_author_id"=>"7403079077"}, {"first_name"=>"Huanzhang", "last_name"=>"Zhu", "scopus_author_id"=>"7404664182"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84869221336", "doi"=>"10.1371/journal.pone.0048832", "pui"=>"366057687", "scopus"=>"2-s2.0-84869221336", "issn"=>"19326203", "pmid"=>"23166597"}, "id"=>"72b290fe-a300-33c6-a2b1-17c89f302176", "abstract"=>"Histone deacetylase (HDAC) inhibitors present an exciting new approach to activate HIV production from latently infected cells to potentially enhance elimination of these cells and achieve a cure. M344, a novel HDAC inhibitor, shows robust activity in a variety of cancer cells and relatively low toxicity compared to trichostatin A (TSA). However, little is known about the effects and action mechanism of M344 in inducing HIV expression in latently infected cells.", "link"=>"http://www.mendeley.com/research/selective-histonedeacetylase-inhibitor-m344-intervenes-hiv1-latency-through-increasing-histone-acety", "reader_count"=>14, "reader_count_by_academic_status"=>{"Researcher"=>5, "Student > Ph. D. Student"=>5, "Student > Bachelor"=>4}, "reader_count_by_user_role"=>{"Researcher"=>5, "Student > Ph. D. Student"=>5, "Student > Bachelor"=>4}, "reader_count_by_subject_area"=>{"Agricultural and Biological Sciences"=>7, "Medicine and Dentistry"=>4, "Neuroscience"=>1, "Chemistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}}, "reader_count_by_country"=>{"Canada"=>1, "United States"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/289772", "https://ndownloader.figshare.com/files/289797", "https://ndownloader.figshare.com/files/289823", "https://ndownloader.figshare.com/files/289849"], "description"=>"<div><h3>Background</h3><p>Histone deacetylase (HDAC) inhibitors present an exciting new approach to activate HIV production from latently infected cells to potentially enhance elimination of these cells and achieve a cure. M344, a novel HDAC inhibitor, shows robust activity in a variety of cancer cells and relatively low toxicity compared to trichostatin A (TSA). However, little is known about the effects and action mechanism of M344 in inducing HIV expression in latently infected cells.</p> <h3>Methodology/Principal Findings</h3><p>Using the Jurkat T cell model of HIV latency, we demonstrate that M344 effectively reactivates HIV-1 gene expression in latently infected cells. Moreover, M344-mediated activation of the latent HIV LTR can be strongly inhibited by a NF-κB inhibitor aspirin. We further show that M344 acts by increasing the acetylation of histone H3 and histone H4 at the nucleosome 1 (nuc-1) site of the HIV-1 long terminal repeat (LTR) and by inducing NF-κB p65 nuclear translocation and direct RelA DNA binding at the nuc-1 region of the HIV-1 LTR. We also found that M344 synergized with prostratin to activate the HIV-1 LTR promoter in latently infected cells.</p> <h3>Conclusions/Significance</h3><p>These results suggest the potential of M344 in anti-latency therapies and an important role for histone modifications and NF-κB transcription factors in regulating HIV-1 LTR gene expression.</p> </div>", "links"=>[], "tags"=>["selective", "histonedeacetylase", "inhibitor", "m344", "intervenes", "hiv-1", "latency", "histone", "acetylation", "activation", "nf-kappab"], "article_id"=>117056, "categories"=>["Molecular Biology", "Cancer", "Genetics"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.s001", "https://dx.doi.org/10.1371/journal.pone.0048832.s002", "https://dx.doi.org/10.1371/journal.pone.0048832.s003", "https://dx.doi.org/10.1371/journal.pone.0048832.s004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Selective_Histonedeacetylase_Inhibitor_M344_Intervenes_in_HIV_1_Latency_through_Increasing_Histone_Acetylation_and_Activation_of_NF_kappaB__/117056", "title"=>"Selective Histonedeacetylase Inhibitor M344 Intervenes in HIV-1 Latency through Increasing Histone Acetylation and Activation of NF-kappaB", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-11-15 01:57:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/541903"], "description"=>"<p>(A) The structure of M344 and TSA. (B) J-Lat clones A7 cells, which have an integrated GFP/Tat construct under control of the HIV-1 LTR, were treated with M344 or TSA at the indicated concentrations for 72 hours. The percentage of cells expressing GFP was measured by flow cytometry, to determine the level of HIV-1 expression. Results are presented as fluorescence histograms. (C) Dose-dependent effects of M344 on HIV-1 production. Data represent the means±standard deviations of three independent experiments.</p>", "links"=>[], "tags"=>["latent", "hiv-1", "latently", "infected", "cells", "m344"], "article_id"=>212388, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Reactivation_of_latent_HIV_1_in_latently_infected_cells_by_M344_and_TSA_/212388", "title"=>"Reactivation of latent HIV-1 in latently infected cells by M344 and TSA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:39:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/542054"], "description"=>"<p>(A) J-Lat clones A7 cells were mock treated or treated with M344 (200 nM) or TSA (100 nM) at the indicated times. GFP expression was monitored in gated live cells at 1, 2, 3, and 4 days by standard flow cytometric techniques. Results are presented as fluorescence histograms. (B) Time-dependent effects of M344 on HIV-1 production. Data are expressed as percentage of cells becoming GFP-positive, and represent the means standard deviations of three independent experiments.</p>", "links"=>[], "tags"=>["reactivation", "latent", "hiv-1", "latently", "infected", "cells", "m344"], "article_id"=>212543, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Kinetic_analysis_of_reactivation_of_latent_HIV_1_in_latently_infected_cells_by_M344_and_TSA_/212543", "title"=>"Kinetic analysis of reactivation of latent HIV-1 in latently infected cells by M344 and TSA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:42:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/542222"], "description"=>"<p>J-Lat clones A7 cells were mock treated or treated with M344 (50 nM), TNF-α (10 ng/ml), 5-Aza (500 nM), prostratin (100 nM), M344/TNF-α, M344/5-Aza or M344/prostratin. The effects of synergistic activation of HIV-1 promoter were determined by quantifying the GFP-positive cells using flow cytometry 72 hours after treatment. Results are presented as fluorescence histograms. Summary of synergistic activation assays are presented as histograms. Data represent the means standard deviations of three independent experiments.</p>", "links"=>[], "tags"=>["activation", "hiv-1", "promoter", "m344", "5-aza", "prostratin", "latently", "infected"], "article_id"=>212701, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Synergistic_activation_of_HIV_1_promoter_by_M344_and_TNF_945_5_Aza_and_prostratin_in_latently_infected_cells_/212701", "title"=>"Synergistic activation of HIV-1 promoter by M344 and TNF-α, 5-Aza and prostratin in latently infected cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:45:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/542327"], "description"=>"<p>293- Human Embryonic Kidney (A), J-Lat clones A7 cells (B) and Jurkat T cells (C) were treated with M344 or TSA at the indicated concentrations for 48 hours, and measured by the MTT method. Results are presented as a percentage of the O.D. (P = 550) of untreated controls subtracted for background. Data represent the means±standard deviations of three independent experiments.</p>", "links"=>[], "tags"=>["viability", "assays", "m344"], "article_id"=>212817, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Summary_of_cell_viability_assays_using_M344_and_TSA_/212817", "title"=>"Summary of cell viability assays using M344 and TSA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:46:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/542426"], "description"=>"<p>(A) Western blot detection of acetylated histone H3 levels in latently infected cells treatment with M344. J-Lat clones A7 cells were mock treated or treated with M344 (100 nM, 200 nM, 400 nM), and cell lysates were harvested after 8 hours. Western blot analysis was performed with antibodies acetylated histone H3. The amount of protein was normalized by comparison to levels of β-actin. (B) Diagram shows the positions of nucleosomes bound to the HIV-1 LTR and the location of primer used for PCR amplification in the ChIP assay. (C)Chromatin fragments from J-Lat clones A7 cells cultured for 4 hours with or without M344 (200 nM) or TSA (200 nM) were immunoprecipitated with antibody to acetylated histones H3 (AcH3) and H4 (AcH4) or control normal rabbit serum (IgG). PCR primers for the LTR promoter were used to amplify the DNA isolated from the immunoprecipitated chromatin as described in Materials and Methods. (D) Each ChIP experiment was repeated three times to confirm reproducibility of results and real-time quantitation of the fold change relative to untreated control is shown.</p>", "links"=>[], "tags"=>["acetylation", "modification", "hiv", "ltr"], "article_id"=>212915, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Histone_acetylation_modification_at_HIV_LTR_promoter_/212915", "title"=>"Histone acetylation modification at HIV LTR promoter.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:48:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/542553"], "description"=>"<p>(A) HDAC6 is present in the nucleus and cytoplasm of J-Lat clones A7 cells. Subcellular localization of HADC6 was assessed by immunofluorescence analysis with a rabbit polyclonal anti-HADC6 IgG antibody and a goat anti-rabbit IgG antibody coupled with Alexa-555 (red color). DAPI staining was used to determine the nuclear region and to assess gross cell morphology. (B) ChIP analysis of HDAC6 occupancy at the HIV-1 LTR promoter in J-Lat clones A7 cells. IGFBP4 promoter was used as a positive-control region of DNA in J-Lat clones A7 cells to verify the ability of HDAC6 antibodies to work in ChIP assays, and rabbit IgG serum was used as a negative control. Chromatin fragments from J-Lat clones A7 cells were immunoprecipitated with antibody to HDAC 6 or control normal rabbit serum (IgG). PCR primers for the LTR promoter or IGFBP4 promoter or IGFBP4 non-targeting DNA were used to amplify the DNA isolated from the immunoprecipitated chromatin as described in Materials and Methods. (C)Each ChIP experiment was repeated three times to confirm reproducibility of results and the values represent the enrichment of LTR DNA over the IgG negative control as determined by quantitative PCR.</p>", "links"=>[], "tags"=>["localization", "hadc6", "j-lat", "clones", "a7", "cells"], "article_id"=>213038, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Subcellular_localization_and_ChIP_analysis_of_HADC6_in_the_J_Lat_clones_A7_cells_model_of_latency_/213038", "title"=>"Subcellular localization and ChIP analysis of HADC6 in the J-Lat clones A7 cells model of latency.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:50:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/542667"], "description"=>"<p>(A) J-Lat clones A7 cells were transfected with HIV1-LTR luc, HIV1-LTRΔκB luc, HIV1-LTRΔAP-1luc, and HIV1-LTRΔSp1luc. At 24 hours posttransfection, the cells were treated or mock treated with M344 (200 nM) or TNF-α (10 ng/ml). Luciferase activity was measured after 24 hours of stimulation. The error bars indicate standard deviation. (B) J-Lat clones A7 cells were pretreated with various concentrations of (0, 2.5, 5 and 10 mM) aspirin for 3 hours and subsequently treated with M344 (100 nM) or TNF-α (10 ng/mL) or prostratin (100 nM) or control medium for 24 hours. The percentage of GFP+ cells (y-axis) in M344 or TNF-α stimulated cells in either the absence or the presence of the chemical inhibitors was measured by flow cytometry. Data represent the means±standard deviations of three independent experiments.</p>", "links"=>[], "tags"=>["activates", "hiv-1", "ltr", "induction"], "article_id"=>213154, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_M344_activates_the_HIV_1_LTR_through_induction_of_NF_954_B_/213154", "title"=>"M344 activates the HIV-1 LTR through induction of NF-κB.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:52:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/542822"], "description"=>"<p>Immunofluorescence analysis of the p65 protein in J-Lat clones A7 cells mock treated or treated with M344, or TNF or TSA for 30 minutes or 2 hours. Subcellular localization of p65 was determined via indirect immunofluorescence employing rabbit polyclonal anti-p65 and goat anti-rabbit antibody coupled to Alexa-555. DAPI staining was used to determine the region of nuclei and to assess gross cell morphology.</p>", "links"=>[], "tags"=>["localization"], "article_id"=>213313, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Subcellular_localization_of_p65_/213313", "title"=>"Subcellular localization of p65.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:55:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/542937"], "description"=>"<p>J-Lat clones A7 were stimulated with M344 (200 nM) or TNF-α(10 ng/ml) for 4 hours, respectively. Chromatin immunoprecipitation assays were performed using anti-p65 or anti-p50 antibodies or rabbit preimmune IgG and probed for the HIV LTR DNA sequences spanning the κB enhancer or for nonspecific control β-actin. Each ChIP experiment was repeated three times to confirm reproducibility of results and real-time quantitation of the fold change relative to untreated control is shown.</p>", "links"=>[], "tags"=>["induces", "rela", "recruitment", "latent", "hiv-1"], "article_id"=>213426, "categories"=>["Virology", "Molecular Biology", "Genetics", "Infectious Diseases"], "users"=>["Hao Ying", "Yuhao Zhang", "Xin Zhou", "Xiying Qu", "Pengfei Wang", "Sijie Liu", "Daru Lu", "Huanzhang Zhu"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048832.g009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_M344_induces_RelA_recruitment_to_the_latent_HIV_1_LTR_/213426", "title"=>"M344 induces RelA recruitment to the latent HIV-1 LTR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-15 00:57:06"}

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Relative Metric

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