Functional Analysis of Conserved Motifs in Influenza Virus PB1 Protein
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{"title"=>"Functional analysis of conserved motifs in influenza virus PB1 protein.", "type"=>"journal", "authors"=>[{"first_name"=>"Caroline", "last_name"=>"Chu", "scopus_author_id"=>"55357546400"}, {"first_name"=>"Shufang", "last_name"=>"Fan", "scopus_author_id"=>"35277167600"}, {"first_name"=>"Chengjun", "last_name"=>"Li", "scopus_author_id"=>"35078736700"}, {"first_name"=>"Catherine", "last_name"=>"Macken", "scopus_author_id"=>"6701809755"}, {"first_name"=>"Jin Hyun", "last_name"=>"Kim", "scopus_author_id"=>"26664082200"}, {"first_name"=>"Masato", "last_name"=>"Hatta", "scopus_author_id"=>"7004521317"}, {"first_name"=>"Gabriele", "last_name"=>"Neumann", "scopus_author_id"=>"7202631658"}, {"first_name"=>"Yoshihiro", "last_name"=>"Kawaoka", "scopus_author_id"=>"26643027000"}], "year"=>2012, "source"=>"PloS one", "identifiers"=>{"issn"=>"19326203", "pui"=>"365627373", "sgr"=>"84866149267", "doi"=>"10.1371/journal.pone.0036113", "scopus"=>"2-s2.0-84866149267", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"22615752"}, "id"=>"db9e0b11-789d-353d-a889-96dc096a7039", "abstract"=>"The influenza virus RNA polymerase complex is a heterotrimer composed of the PB1, PB2, and PA subunits. PB1, the catalytic core and structural backbone of the polymerase, possesses four highly conserved amino acid motifs that are present among all viral RNA-dependent RNA polymerases. A previous study demonstrated the importance of several of these conserved amino acids in PB1 for influenza polymerase activity through mutational analysis. However, a small number of viruses isolated in nature possesses non-consensus amino acids in one of the four motifs, most of which have not been tested for their replicative ability. Here, we assessed the transcription/replication activities of 25 selected PB1 mutations found in natural isolates by using minireplicon assays in human and avian cells. Most of the mutations tested significantly reduced polymerase activity. One exception was mutation K480R, observed in several pandemic (H1N1) 2009 viruses, which slightly increased polymerase activity relative to wild-type. However, in the background of the pandemic A/California/04/2009 (H1N1) virus, this mutation did not affect virus titers in cell culture. Our results further demonstrate the functional importance of the four conserved PB1 motifs in influenza virus transcription/replication. The finding of natural isolates with non-consensus PB1 motifs that are nonfunctional in minireplicon assays suggests compensatory mutations and/or mixed infections which may have 'rescued' the inactive PB1 protein.", "link"=>"http://www.mendeley.com/research/functional-analysis-conserved-motifs-influenza-virus-pb1-protein", "reader_count"=>31, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Researcher"=>4, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>2, "Student > Master"=>5, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Researcher"=>4, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>2, "Student > Master"=>5, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>20, "Medicine and Dentistry"=>1, "Veterinary Science and Veterinary Medicine"=>2, "Business, Management and Accounting"=>1, "Social Sciences"=>1, "Immunology and Microbiology"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>20}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Denmark"=>1, "India"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/638447"], "description"=>"*<p>Percentage of total changes.</p>", "links"=>[], "tags"=>["pb1", "proteins", "mutations", "conserved", "motifs"], "article_id"=>308928, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.t002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Avian_virus_PB1_proteins_with_mutations_in_the_conserved_motifs_I_8211_IV_/308928", "title"=>"Avian virus PB1 proteins with mutations in the conserved motifs I – IV.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:11:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/638340"], "description"=>"<p>Wild-type A/California/04/09 virus and recombinant virus possessing the K480R mutation were used for infection of human Calu-3 cells (MOI = 0.001). Supernatants were collected at specified time points following infection, and virus titers were quantified by plaque assays in MDCK cells. Results shown are averages of three plaque assays.</p>", "links"=>[], "tags"=>["characteristics", "wild-type", "possessing", "pb1", "k480r"], "article_id"=>308819, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.g005", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Growth_characteristics_of_wild_type_virus_and_virus_possessing_the_PB1_K480R_mutation_/308819", "title"=>"Growth characteristics of wild-type virus and virus possessing the PB1 K480R mutation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:10:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/638259"], "description"=>"<p>The effects of PB1 mutation K480R were evaluated in the background of the A/California/04/09 (H1N1) virus replication complex in human 293T and chicken DF-1 cells. The experiments were carried out as described in the legend to <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036113#pone-0036113-g001\" target=\"_blank\">Figure 1</a>. Luciferase activity levels were normalized by the <i>Renilla</i> internal control and expressed as a percentage relative to wild-type PB1 activity in the particular cell line. *P<0.07 by Student’s t-test.</p>", "links"=>[], "tags"=>["pb1", "mutation", "k480r", "influenza", "polymerase", "avian"], "article_id"=>308745, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.g004", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_PB1_mutation_K480R_on_influenza_polymerase_activity_in_human_and_avian_cells_/308745", "title"=>"Effects of PB1 mutation K480R on influenza polymerase activity in human and avian cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:10:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/638114"], "description"=>"<p>293T cells were transfected with plasmids expressing PB2, PA, NP, and wild-type or mutant PB1 protein, along with a plasmid transcribing a viral RNA. The levels of vRNA, cRNA, and mRNA were assessed by quantitative RT-PCR and compared to those obtained with wild-type PB1 protein. The results shown represent the average levels of RNAs ± standard deviations from a representative experiment carried out in triplicate. As a negative control (Mock), the PB1 protein expression plasmid was omitted.</p>", "links"=>[], "tags"=>["mrna", "levels", "wild-type", "mutant", "pb1"], "article_id"=>308592, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.g002", "stats"=>{"downloads"=>2, "page_views"=>27, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantitative_analysis_of_vRNA_cRNA_and_mRNA_levels_for_wild_type_and_mutant_PB1_proteins_/308592", "title"=>"Quantitative analysis of vRNA, cRNA, and mRNA levels for wild-type and mutant PB1 proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:09:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/638489"], "description"=>"1<p>The indicated mutations were introduced into the PB1 protein of A/bar-headed goose/Qinghai/1/2005 (H5N1; BHG) virus. Polymerase activity in human 293T and avian DF-1 cells is indicated as percentage relative to wild-type PB1 activity.</p>2<p>For some of these isolates, different sequences are reported in the Los Alamos National Laboratory and NIH NCBI databases.</p>", "links"=>[], "tags"=>["conserved", "motifs", "iv", "pb1", "replicative"], "article_id"=>308974, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.t003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mutations_in_the_conserved_motifs_I_8211_IV_of_the_PB1_protein_and_their_effects_on_replicative_ability_in_human_and_chicken_cells_/308974", "title"=>"Mutations in the conserved motifs I – IV of the PB1 protein and their effects on replicative ability in human and chicken cells.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:11:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/638182"], "description"=>"<p>293T cells were transfected with plasmids expressing wild-type or mutant PB1 protein. Western blot analysis was carried out with antibodies to PB1 and beta-actin (control).</p>", "links"=>[], "tags"=>["blot", "wild-type", "mutant", "pb1"], "article_id"=>308668, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Western_blot_analysis_of_wild_type_and_mutant_PB1_proteins_/308668", "title"=>"Western blot analysis of wild-type and mutant PB1 proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:09:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/638015"], "description"=>"<p>(<b>A</b>) Motif I; (<b>B</b>) Motif II; (<b>C</b>) Motif III; (<b>D</b>) Motif IV. Consensus sequences are listed by motif with numbers at each end indicating the position of the first and last amino acid residue. Invariant amino acids are underlined. Selected mutations found in the four conserved motifs of natural isolates (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036113#pone-0036113-t003\" target=\"_blank\"><b>Table 3</b></a>) were introduced into the PB1 protein of A/bar-headed goose/Qinghai/1/2005 (H5N1) virus. The polymerase activity of each PB1 mutant was assessed in human 293T cells by quantifying luciferase activity in the cell lysates of cells transfected with PB1- (wild type or mutant), PB2-, PA- and NP-protein expression plasmids along with a reporter plasmid expressing an influenza virus-like RNA construct for luciferase. Luciferase activity levels were normalized by the <i>Renilla</i> internal control and expressed as a percentage relative to wild-type PB1 activity set as 100%. Data values are averaged over three independent transfection experiments where each experiment was run in triplicate. Error bars represent one standard deviation.</p>", "links"=>[], "tags"=>["mutations", "conserved", "motifs", "pb1", "influenza", "polymerase"], "article_id"=>308490, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.g001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_mutations_in_the_four_conserved_motifs_of_PB1_on_influenza_polymerase_activity_/308490", "title"=>"Effects of mutations in the four conserved motifs of PB1 on influenza polymerase activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:08:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/638401"], "description"=>"*<p>Percentage of total changes.</p>", "links"=>[], "tags"=>["pb1", "proteins", "mutations", "conserve", "motifs"], "article_id"=>308883, "categories"=>["Biochemistry", "Biological Sciences", "Physics", "Infectious Diseases", "Biophysics"], "users"=>["Caroline Chu", "Shufang Fan", "Chengjun Li", "Catherine Macken", "Jin Hyun Kim", "Masato Hatta", "Gabriele Neumann", "Yoshihiro Kawaoka"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036113.t001", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Influenza_A_virus_PB1_proteins_with_mutations_in_the_conserve_motifs_I_8211_IV_/308883", "title"=>"Influenza A virus PB1 proteins with mutations in the conserve motifs I – IV.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:10:55"}

PMC Usage Stats | Further Information

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Relative Metric

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