Transcriptional and Proteomic Analysis of the Aspergillus fumigatus ΔprtT Protease-Deficient Mutant
Publication Date
April 13, 2012
Journal
PLOS ONE
Authors
Shelly Hagag, Paula Kubitschek Barreira, Gabriela W. P. Neves, David Amar, et al
Volume
7
Issue
4
Pages
e33604
DOI
https://dx.plos.org/10.1371/journal.pone.0033604
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0033604
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/22514608
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3326020
Europe PMC
http://europepmc.org/abstract/MED/22514608
Web of Science
000305341600013
Scopus
84859724704
Mendeley
http://www.mendeley.com/research/transcriptional-proteomic-analysis-aspergillus-fumigatus-%CE%B4prtt-proteasedeficient-mutant
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Mendeley 3327 Apr 14:13 UTC

Mendeley | Further Information

{"title"=>"Transcriptional and proteomic analysis of the aspergillus fumigatus δprtt protease-deficient mutant", "type"=>"journal", "authors"=>[{"first_name"=>"Shelly", "last_name"=>"Hagag", "scopus_author_id"=>"8858559500"}, {"first_name"=>"Paula", "last_name"=>"Kubitschek-Barreira", "scopus_author_id"=>"55183909600"}, {"first_name"=>"Gabriela W.P.", "last_name"=>"Neves", "scopus_author_id"=>"55183966900"}, {"first_name"=>"David", "last_name"=>"Amar", "scopus_author_id"=>"37070387400"}, {"first_name"=>"William", "last_name"=>"Nierman", "scopus_author_id"=>"7003633617"}, {"first_name"=>"Itamar", "last_name"=>"Shalit", "scopus_author_id"=>"7004096224"}, {"first_name"=>"Ron", "last_name"=>"Shamir", "scopus_author_id"=>"7005411902"}, {"first_name"=>"Leila", "last_name"=>"Lopes-Bezerra", "scopus_author_id"=>"6602403549"}, {"first_name"=>"Nir", "last_name"=>"Osherov", "scopus_author_id"=>"6701736159"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"22514608", "sgr"=>"84859724704", "doi"=>"10.1371/journal.pone.0033604", "scopus"=>"2-s2.0-84859724704", "pui"=>"364617706", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "issn"=>"19326203"}, "id"=>"133a820f-33b2-39fe-ab2b-a7c88a41bd63", "abstract"=>"Aspergillus fumigatus is the most common opportunistic mold pathogen of humans, infecting immunocompromised patients. The fungus invades the lungs and other organs, causing severe damage. Penetration of the pulmonary epithelium is a key step in the infectious process. A. fumigatus produces extracellular proteases to degrade the host structural barriers. The A. fumigatus transcription factor PrtT controls the expression of multiple secreted proteases. PrtT shows similarity to the fungal Gal4-type Zn(2)-Cys(6) DNA-binding domain of several transcription factors. In this work, we further investigate the function of this transcription factor by performing a transcriptional and a proteomic analysis of the ΔprtT mutant. Unexpectedly, microarray analysis revealed that in addition to the expected decrease in protease expression, expression of genes involved in iron uptake and ergosterol synthesis was dramatically decreased in the ΔprtT mutant. A second finding of interest is that deletion of prtT resulted in the upregulation of four secondary metabolite clusters, including genes for the biosynthesis of toxic pseurotin A. Proteomic analysis identified reduced levels of three secreted proteases (ALP1 protease, TppA, AFUA_2G01250) and increased levels of three secreted polysaccharide-degrading enzymes in the ΔprtT mutant possibly in response to its inability to derive sufficient nourishment from protein breakdown. This report highlights the complexity of gene regulation by PrtT, and suggests a potential novel link between the regulation of protease secretion and the control of iron uptake, ergosterol biosynthesis and secondary metabolite production in A. fumigatus.", "link"=>"http://www.mendeley.com/research/transcriptional-proteomic-analysis-aspergillus-fumigatus-%CE%B4prtt-proteasedeficient-mutant", "reader_count"=>32, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>7, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Master"=>10, "Other"=>2, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>7, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Master"=>10, "Other"=>2, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>22, "Medicine and Dentistry"=>1, "Chemistry"=>1, "Computer Science"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>22}, "Computer Science"=>{"Computer Science"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}}, "reader_count_by_country"=>{"Brazil"=>1, "Germany"=>1}, "group_count"=>1}
Scopus 1714 Feb 20:15 UTC

Scopus | Further Information

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Figshare 4602 Apr 21:01 UTC

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  • {"files"=>["https://ndownloader.figshare.com/files/653415"], "description"=>"1<p>genes also identified by Wartenberg ,et al. (43)</p>", "links"=>[], "tags"=>["proteins"], "article_id"=>323904, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.t004", "stats"=>{"downloads"=>2, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Secretome_proteins_identified_by_2D_8722_DIGE_/323904", "title"=>"Secretome proteins identified by 2D−DIGE.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-04-13 01:05:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/653492"], "description"=>"1<p>Top 10 genes with the highest fold change are shown in every category.</p>2<p>Gene located in a gene cluster.</p>", "links"=>[], "tags"=>["enriched", "upregulated", "classes", "mutant"], "article_id"=>323976, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.t002", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Selected_enriched_upregulated_gene_classes_in_the_916_prtT_mutant_vs_W_/323976", "title"=>"Selected enriched upregulated gene classes in the <i>ΔprtT</i> mutant vs. W.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-04-13 01:06:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/653195"], "description"=>"<p>Expression rates were normalized relative to mRNA levels of the β-tubulin-encoding gene (AFUA_1G10910) and set arbitrarily to 1 for the WT strain grown in 1% SM medium. Values are given in relative quantity of template compared to the original sample (RQ). RQ values were calculated by use of the equation: RQ  =  2<sup>–ΔΔCT</sup>, with ΔΔCT ± SD and ΔCT ± SDs. −Fe  =  growth medium lacking iron and containing 1 mM ferrozine. +Vori  =  growth medium supplemented with a sub-inhibitory concentration (0.125 µg/ml) of voriconazole. The experiment was repeated three times with similar results. Graphs show a representative experiment.</p>", "links"=>[], "tags"=>["transcripts", "levels", "genes", "showed", "differential", "microarray", "encoding", "transcription", "factors"], "article_id"=>323669, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_qRT_PCR_evaluation_of_transcripts_levels_from_genes_that_showed_a_differential_expression_in_the_microarray_A_representative_genes_MirB_SidA_Erg3_Erg11_and_Alp1_and_B_the_genes_HapX_SreA_AcuM_and_SrbA_encoding_transcription_factors_that_regulate_iron_upt/323669", "title"=>"qRT-PCR evaluation of transcripts levels from genes that showed a differential expression in the microarray (A) representative genes (<i>MirB, SidA, Erg3, Erg11</i> and <i>Alp1</i>) and (B) the genes <i>HapX</i>, <i>SreA, AcuM</i> and <i>SrbA</i> encoding transcription factors that regulate iron uptake.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-04-13 01:01:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/653043"], "description"=>"<p>Black bars are the iron uptake genes, white bars are genes with various other annotations.</p>", "links"=>[], "tags"=>["50", "downregulated", "genes", "plotted", "ascending"], "article_id"=>323523, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Top_50_downregulated_genes_plotted_in_ascending_order_by_fold_change_/323523", "title"=>"Top 50 downregulated genes plotted in ascending order by fold change.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-04-13 00:58:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/653343"], "description"=>"<p>DIGE image overlay of WT (Cy3 label in green) and <i>ΔprtT</i> (Cy5 label in red) secretomes. Yellow spots indicate proteins detected in both strains and identified proteins are indicated by their accession number (SwissProt/UniProtKB database). Detected protein isoforms are represented with Arabic numerals.</p>", "links"=>[], "tags"=>["dige", "comparing", "secretomes", "wt"], "article_id"=>323828, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.g004", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_2_D_DIGE_comparing_the_secretomes_of_WT_and_916_prtT_strains_/323828", "title"=>"2-D DIGE comparing the secretomes of WT and <i>ΔprtT</i> strains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-04-13 01:03:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/653453"], "description"=>"<p>1Top five genes with the highest fold change are shown in each category.</p>2<p>Ergosterol biosynthesis pathway.</p>3<p>Involved in amino acid catabolism.</p>", "links"=>[], "tags"=>["enriched", "downregulated", "classes", "mutant"], "article_id"=>323938, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.t001", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Selected_enriched_downregulated_gene_classes_in_the_916_prtT_mutant_vs_WT_1_/323938", "title"=>"Selected enriched downregulated gene classes in the <i>ΔprtT</i> mutant vs. WT<sup>1</sup>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-04-13 01:05:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/653118"], "description"=>"<p>(A) Growth in iron-limited media: WT and <i>ΔprtT</i> mutant were grown in iron-lacking 0.1% SM agar and supplemented with different concentrations of the ferrous iron chelator ferrozine. Colony diameter was measured after 48 h at 37°C. Similar results were seen with 1% SM and using liquid medium as well (data not shown). (B) Evaluation of siderophore production using the CAS assay. WT and <i>ΔprtT</i> conidia were grown either in liquid medium containing iron (indicated by +Fe) or on medium without iron (indicated by −Fe). Culture supernatants were collected and added to wells made in the CAS agar plates. The presence of a halo around the well indicates siderophore production. Non-inoculated growth media were also applied to wells on the CAS plate as a negative control (Medium) (C). Sensitivity to voriconazole. WT and <i>ΔprtT</i> mutant were grown on 1% SM agar plates supplemented with different concentrations of voriconazole. Colony diameter was measured after 48 h at 37°C.</p>", "links"=>[], "tags"=>["genetics and genomics", "Infectious diseases", "microbiology", "Computational biology", "Biochemistry"], "article_id"=>323599, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phenotypic_analysis_of_the_916_prtT_mutant_/323599", "title"=>"Phenotypic analysis of the <i>ΔprtT</i> mutant.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-04-13 00:59:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/653535"], "description"=>"<p>Significantly enriched physical clusters</p>", "links"=>[], "tags"=>["enriched"], "article_id"=>324022, "categories"=>["Biochemistry", "Microbiology", "Biological Sciences", "Genetics", "Infectious Diseases"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0033604.t003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Significantly_enriched_physical_clusters_/324022", "title"=>"Significantly enriched physical clusters", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-04-13 01:07:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/336064", "https://ndownloader.figshare.com/files/336094"], "description"=>"<div><p><em>Aspergillus fumigatus</em> is the most common opportunistic mold pathogen of humans, infecting immunocompromised patients. The fungus invades the lungs and other organs, causing severe damage. Penetration of the pulmonary epithelium is a key step in the infectious process. <em>A. fumigatus</em> produces extracellular proteases to degrade the host structural barriers. The <em>A. fumigatus</em> transcription factor PrtT controls the expression of multiple secreted proteases. PrtT shows similarity to the fungal Gal4-type Zn(2)-Cys(6) DNA-binding domain of several transcription factors. In this work, we further investigate the function of this transcription factor by performing a transcriptional and a proteomic analysis of the <em>ΔprtT</em> mutant. Unexpectedly, microarray analysis revealed that in addition to the expected decrease in protease expression, expression of genes involved in iron uptake and ergosterol synthesis was dramatically decreased in the <em>ΔprtT</em> mutant. A second finding of interest is that deletion of <em>prtT</em> resulted in the upregulation of four secondary metabolite clusters, including genes for the biosynthesis of toxic pseurotin A. Proteomic analysis identified reduced levels of three secreted proteases (ALP1 protease, TppA, AFUA_2G01250) and increased levels of three secreted polysaccharide-degrading enzymes in the <em>ΔprtT</em> mutant possibly in response to its inability to derive sufficient nourishment from protein breakdown. This report highlights the complexity of gene regulation by PrtT, and suggests a potential novel link between the regulation of protease secretion and the control of iron uptake, ergosterol biosynthesis and secondary metabolite production in <em>A. fumigatus</em>.</p> </div>", "links"=>[], "tags"=>["transcriptional", "proteomic", "fumigatus", "protease-deficient", "mutant"], "article_id"=>126399, "categories"=>["Biochemistry", "Cancer", "Biological Sciences", "Genetics", "Microbiology"], "users"=>["Shelly Hagag", "Paula Kubitschek-Barreira", "Gabriela W. P. Neves", "David Amar", "William Nierman", "Itamar Shalit", "Ron Shamir", "Leila Lopes-Bezerra", "Nir Osherov"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0033604.s001", "https://dx.doi.org/10.1371/journal.pone.0033604.s002"], "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Transcriptional_and_Proteomic_Analysis_of_the_Aspergillus_fumigatus_prtT_Protease_Deficient_Mutant/126399", "title"=>"Transcriptional and Proteomic Analysis of the <em>Aspergillus fumigatus ΔprtT</em> Protease-Deficient Mutant", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-04-13 01:46:39"}
PMC Usage Stats 111417 Feb 17:00 UTC

PMC Usage Stats | Further Information

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Relative Metric 15173105 Apr 19:01 UTC

Relative Metric

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F1000Prime 124 Feb 12:16 UTC

F1000Prime | Further Information

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