MiR-125b Reduces Porcine Reproductive and Respiratory Syndrome Virus Replication by Negatively Regulating the NF-κB Pathway
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{"title"=>"MiR-125b Reduces Porcine Reproductive and Respiratory Syndrome Virus Replication by Negatively Regulating the NF-κB Pathway", "type"=>"journal", "authors"=>[{"first_name"=>"Dang", "last_name"=>"Wang", "scopus_author_id"=>"36060664000"}, {"first_name"=>"Lu", "last_name"=>"Cao", "scopus_author_id"=>"39860906400"}, {"first_name"=>"Zheng", "last_name"=>"Xu", "scopus_author_id"=>"57198998272"}, {"first_name"=>"Liurong", "last_name"=>"Fang", "scopus_author_id"=>"7402470374"}, {"first_name"=>"Yao", "last_name"=>"Zhong", "scopus_author_id"=>"54392097300"}, {"first_name"=>"Quangang", "last_name"=>"Chen", "scopus_author_id"=>"38460892900"}, {"first_name"=>"Rui", "last_name"=>"Luo", "scopus_author_id"=>"35230419100"}, {"first_name"=>"Huanchun", "last_name"=>"Chen", "scopus_author_id"=>"7501622358"}, {"first_name"=>"Kui", "last_name"=>"Li", "scopus_author_id"=>"55733139900"}, {"first_name"=>"Shaobo", "last_name"=>"Xiao", "scopus_author_id"=>"7402022567"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84873586638", "pui"=>"368311202", "doi"=>"10.1371/journal.pone.0055838", "isbn"=>"1932-6203", "sgr"=>"84873586638", "pmid"=>"23409058"}, "id"=>"a168c7f4-0284-364c-9e4f-c746db59ba62", "abstract"=>"Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus that has been devastating the swine industry worldwide since the late 1980s. To investigate the impact of cellular microRNAs (miRNAs) on the replication of PRRSV, we screened 10 highly conserved miRNAs implicated in innate immunity or antiviral function and identified miR-125b as an inhibitor of PRRSV replication. Virus titer and western blot assays demonstrated that miR-125b reduced PRRSV replication and viral gene expression in a dose-dependent manner in both MARC-145 cell line and primary porcine alveolar macrophages. Mechanistically, miR-125b did not target the PRRSV genome. Rather, it inhibited activation of NF-κB, which we found to be required for PRRSV replication. PRRSV, in turn, down-regulated miR-125b expression post-infection to promote viral replication. Collectively, miR-125b is an antiviral host factor against PRRSV, but it is subject to manipulation by PRRSV. Our study reveals an example of manipulation of a cellular miRNA by an arterivirus to re-orchestrate host gene expression for viral propagation and sheds new light on targeting host factors to develop effective control measures for PRRS.", "link"=>"http://www.mendeley.com/research/mir125b-reduces-porcine-reproductive-respiratory-syndrome-virus-replication-negatively-regulating-nf-3", "reader_count"=>5, "reader_count_by_academic_status"=>{"Researcher"=>1, "Student > Master"=>1, "Other"=>3}, "reader_count_by_user_role"=>{"Researcher"=>1, "Student > Master"=>1, "Other"=>3}, "reader_count_by_subject_area"=>{"Agricultural and Biological Sciences"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Unspecified"=>3}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>1}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/483760", "https://ndownloader.figshare.com/files/483765"], "description"=>"<div><p>Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus that has been devastating the swine industry worldwide since the late 1980s. To investigate the impact of cellular microRNAs (miRNAs) on the replication of PRRSV, we screened 10 highly conserved miRNAs implicated in innate immunity or antiviral function and identified miR-125b as an inhibitor of PRRSV replication. Virus titer and western blot assays demonstrated that miR-125b reduced PRRSV replication and viral gene expression in a dose-dependent manner in both MARC-145 cell line and primary porcine alveolar macrophages. Mechanistically, miR-125b did not target the PRRSV genome. Rather, it inhibited activation of NF-κB, which we found to be required for PRRSV replication. PRRSV, in turn, down-regulated miR-125b expression post-infection to promote viral replication. Collectively, miR-125b is an antiviral host factor against PRRSV, but it is subject to manipulation by PRRSV. Our study reveals an example of manipulation of a cellular miRNA by an arterivirus to re-orchestrate host gene expression for viral propagation and sheds new light on targeting host factors to develop effective control measures for PRRS.</p> </div>", "links"=>[], "tags"=>["mir-125b", "reduces", "porcine", "reproductive", "respiratory", "replication", "negatively", "regulating", "pathway"], "article_id"=>156200, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0055838.s001", "https://dx.doi.org/10.1371/journal.pone.0055838.s002"], "stats"=>{"downloads"=>7, "page_views"=>53, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/MiR_125b_Reduces_Porcine_Reproductive_and_Respiratory_Syndrome_Virus_Replication_by_Negatively_Regulating_the_NF_B_Pathway__/156200", "title"=>"MiR-125b Reduces Porcine Reproductive and Respiratory Syndrome Virus Replication by Negatively Regulating the NF-κB Pathway", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-02-07 01:43:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/493402"], "description"=>"<p>(A) Overexpression of the NF-κB p65 subunit promotes PRRSV replication and partially antagonizes miR-125b’s effect on PRRSV. MARC-145 cells were cotransfected with a control vector or vector encoding p65 (1.0 µg) and 60 nM of miR-125b mimic or inhibitor. The transfected cells were infected with PRRSV WUH3 strain (MOI = 0.01) 24 h later. Cells were collected at 48 h post-infection for plaque assay on MARC-145 cells. Virus titers were expressed as PFU/mL. Representative plaque results from three independent experiments are shown in left panel and the average results are illustrated on the right. **P<0.01 and *P<0.05 as compared with cells transfected with the control vector. (B, C) Pretreatment with the NF-κB inhibitor BAY11-7082 reduces PRRSV replication in MARC-145 cells (2.5 µM, 5.0 µM and 10µM of BAY11-7082, panel B) and PAMs (5 µM, panel C). Cells were pretreated with BAY11-7082 for 1 h prior to PRRSV infection. At 48 h post-infection, cells were collected and virus titers were determined by plaque assay on MARC-145 cells. (D) The time-course expression of miR-125b after PRRSV infection. MARC-145 cells infected with PRRSV at a MOI of 0.1 were collected at the indicated time points and qRT-PCR analysis was performed to detect miR-125b expression. The miR-125b expression level at 6 h in mock-infected cells was used as the baseline (1.0) for comparison.</p>", "links"=>[], "tags"=>["inter-relationship", "activation", "prrsv"], "article_id"=>163913, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g006", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_inter_relationship_among_miR_125b_NF_954_B_activation_and_PRRSV_replication_/163913", "title"=>"The inter-relationship among miR-125b, NF-κB activation and PRRSV replication.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 01:05:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/493180"], "description"=>"<p>(A) MARC-145 cells were co-transfected with IFN-β-Luc, pRL-TK, and 30 nM of miR-125b mimic or NC mimic. 24 h post-transfection, selected wells were transfected with poly(I:C) (2 µg/mL). After another 24 h, cells were lysed for dual-luciferase assay. (B) MARC-145 cells were transfected with miR-125b mimic (30 nM), NC mimic (30 nM), poly(I:C) (1 µg/mL), or mock-transfected. At 24 h post transfection, cells were infected with VSV-GFP at MOI of 0.0001. Cells were fixed at 24 h post-infection and cellular nuclei were counterstained with 1 µg/mL of DAPI. Fluorescence was observed under an LSM-510 Meta confocal fluorescence microscope (Zeiss).</p>", "links"=>[], "tags"=>["induce", "ifn"], "article_id"=>163691, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_125b_does_not_induce_the_IFN_pathway_/163691", "title"=>"miR-125b does not induce the IFN pathway.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 01:01:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/493301"], "description"=>"<p>(A) Alignment of miR-125b sequence with the 3′UTR of human (GenBank accession no. NM_001001349), monkey (GenBank accession no. XM_001093984) and predicted porcine (GenBank accession no. AK348039) κB-Ras2. Solid lines indicate Watson-Crick base pairs. Dotted line indicates GU wobble pairs. (B) MARC-145 cells were transfected with miR-125b mimic or NC mimic, and analyzed for κB-Ras2 mRNA expression 48 h later by qPCR. *p<0.05 <i>vs</i> NC mimic. (C) MARC-145 cells were co-transfected with pNF-κB-Luc, pRL-TK, and the indicated dose of miR-125b mimic or NC mimic. At 48 h post-transfection, cells were lysed for dual-luciferase assay. (D, E) miR-125b reduces PRRSV-induced NF-κB activation. MARC-145 cells were co-transfected with 0.1 µg of pNF-κB-Luc, 0.05 µg of pRL-TK, and 60 nM of miR-125b mimic (D) or inhibitor (E), followed by PRRSV infection 24 h later. Cells were lysed at 48 h post-infection for dual-luciferase assay. **P<0.01 as compared with NC mimic or inhibitor.</p>", "links"=>[], "tags"=>["negatively", "regulates"], "article_id"=>163816, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g005", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_125b_negatively_regulates_NF_954_B_activation_/163816", "title"=>"miR-125b negatively regulates NF-κB activation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 01:03:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/492876"], "description"=>"<p>(A) Overexpression of miR-125b mimic reduced PRRSV replication in a dose-dependent manner. MARC-145 cells were transfected with miR-125b mimic or a control mimic (NC) at the indicated dose (30, 60, 120 nM), followed by PRRSV infection (MOI = 0.1). The infected cells were collected 48 h later for plaque assays on MARC-145 cells. The plaque results shown on the left were representative of three independent experiments (right). “**” denotes significant difference (P<0.01). (B) Overexpression of miR-125b mimic reduced the expression of PRRSV Nsp2 protein. MARC-145 cells were transfected with miR-125b mimic or the control mimic prior to PRRSV infection. Cells were collected at 30 h post-infection for western blot analysis of nsp2 expression using a specific monoclonal antibody against Nsp2 as the primary antibody. Expression of β-actin was analyzed as a loading control (left). The nsp2 expression levels were quantitated by densitometry using a Gel-Pro analyzer (Right). Data shown were representative of three independent experiments. (C) Overexpression of miR-125b mimic reduced PRRSV replication in PAMs. PAMs were transfected with miR-125b mimic or negative control (60 nM), followed by PRRSV infection (MOI = 0.1). At 48 h post-infection, cells were collected for plaque assay. Representative plaque results were shown in left panel and the data from three independent experiments were plotted in the right panel. (D) Immunofluorescence staining confirms the reduction effect of miR-125b on PRRSV replication in PAMs. The PAMs were transfected with miR-125b mimic or negative control (60 nM), followed by PRRSV infection (MOI = 0.1). Cells were fixed at 24 h post-infection and immunostained with a fluorescein isothiocyanate (FITC)-conjugated monoclonal antibody against the PRRSV N protein. Cellular nuclei were counterstained with 1 µg/mL of DAPI. Fluorescence was observed under an LSM-510 Meta confocal fluorescence microscope (Zeiss).</p>", "links"=>[], "tags"=>["prrsv", "replication", "marc-145", "cells", "porcine", "alveolar", "macrophages"], "article_id"=>163380, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g002", "stats"=>{"downloads"=>3, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_125b_reduces_PRRSV_replication_in_MARC_145_cells_and_porcine_alveolar_macrophages_PAMs_/163380", "title"=>"miR-125b reduces PRRSV replication in MARC-145 cells and porcine alveolar macrophages (PAMs).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:56:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/492968"], "description"=>"<p>(A) Schematic representation of the PRRSV genome. Viral cDNA fragments used for constructing the pMIR-REPORT-derived luciferase reporters are as indicated. (B) MARC-145 cells were co-transfected with 0.1 µg of indicated constructing luciferase reporter, 0.05 µg of pRL-TK, and 30 nM of miR-125b mimic or negative control of mimic. At 24 h post-transfection, cells were lysed for dual-luciferase assay. The relative luciferase activities (miR-125b/NC) refer to fold change in luciferase activity in miR-125b mimic-transfected cells relative to respective NC mimic-transfected controls.</p>", "links"=>[], "tags"=>["prrsv"], "article_id"=>163479, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g003", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_125b_does_not_directly_target_the_PRRSV_genome_/163479", "title"=>"miR-125b does not directly target the PRRSV genome.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:57:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/493479"], "description"=>"<p>PRRSV infection down-regulates the expression of miR-125b, which relieves the stabilizing effect on κB-Ras2 mRNA, resulting in subsequent NF-κB activation. The activated NF-κB promotes PRRSV replication.</p>", "links"=>[], "tags"=>["inter-relationship", "activation", "prrsv"], "article_id"=>163994, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g007", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_proposed_model_for_the_inter_relationship_among_miR_125b_NF_954_B_activation_and_PRRSV_replication_/163994", "title"=>"The proposed model for the inter-relationship among miR-125b, NF-κB activation and PRRSV replication.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 01:06:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/492720"], "description"=>"<p>Confluent MARC-145 cells were transfected with the indicated mimics (A) or inhibitors (B) of the indicated miRNAs. 24 h later, cells were infected with PRRSV strain WUH3 at a multiplicity of infection (MOI) of 0.1. Cells were collected 48 h later for plaque assays on MARC-145 cells. Virus titers were expressed as plaque forming units (PFU)/mL.</p>", "links"=>[], "tags"=>["identifies", "mir-125b", "inhibitor", "porcine", "reproductive", "respiratory"], "article_id"=>163230, "categories"=>["Virology", "Genetics", "Microbiology"], "users"=>["Dang Wang", "Lu Cao", "Zheng Xu", "Liurong Fang", "Yao Zhong", "Quangang Chen", "Rui Luo", "Huanchun Chen", "Kui Li", "Shaobo Xiao"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055838.g001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MicroRNA_miRNA_screening_identifies_miR_125b_as_an_inhibitor_of_porcine_reproductive_and_respiratory_syndrome_virus_PRRSV_replication_/163230", "title"=>"MicroRNA (miRNA) screening identifies miR-125b as an inhibitor of porcine reproductive and respiratory syndrome virus (PRRSV) replication.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-07 00:53:50"}

PMC Usage Stats | Further Information

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Relative Metric

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