The Plasmodium berghei Ca2+/H+ Exchanger, PbCAX, Is Essential for Tolerance to Environmental Ca2+ during Sexual Development
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{"title"=>"The Plasmodium berghei Ca2+/H+ Exchanger, PbCAX, Is Essential for Tolerance to Environmental Ca2+ during Sexual Development", "type"=>"journal", "authors"=>[{"first_name"=>"David S.", "last_name"=>"Guttery", "scopus_author_id"=>"31267552700"}, {"first_name"=>"Jon K.", "last_name"=>"Pittman", "scopus_author_id"=>"7102551810"}, {"first_name"=>"Karine", "last_name"=>"Frénal", "scopus_author_id"=>"6505622506"}, {"first_name"=>"Benoit", "last_name"=>"Poulin", "scopus_author_id"=>"7003681920"}, {"first_name"=>"Leon R.", "last_name"=>"McFarlane", "scopus_author_id"=>"55808555700"}, {"first_name"=>"Ksenija", "last_name"=>"Slavic", "scopus_author_id"=>"28167771800"}, {"first_name"=>"Sally P.", "last_name"=>"Wheatley", "scopus_author_id"=>"7004330121"}, {"first_name"=>"Dominique", "last_name"=>"Soldati-Favre", "scopus_author_id"=>"8235046700"}, {"first_name"=>"Sanjeev", "last_name"=>"Krishna", "scopus_author_id"=>"7102243526"}, {"first_name"=>"Rita", "last_name"=>"Tewari", "scopus_author_id"=>"8343732400"}, {"first_name"=>"Henry M.", "last_name"=>"Staines", "scopus_author_id"=>"7004489641"}], "year"=>2013, "source"=>"PLoS Pathogens", "identifiers"=>{"issn"=>"15537366", "isbn"=>"1553-7374 (Electronic)\\r1553-7366 (Linking)", "pui"=>"369459540", "sgr"=>"84880949759", "doi"=>"10.1371/journal.ppat.1003191", "scopus"=>"2-s2.0-84880949759", "pmid"=>"23468629"}, "id"=>"b283e146-e0df-37a0-bb9f-0c911e6ba491", "abstract"=>"Ca(2+) contributes to a myriad of important cellular processes in all organisms, including the apicomplexans, Plasmodium and Toxoplasma. Due to its varied and essential roles, free Ca(2+) is tightly regulated by complex mechanisms. These mechanisms are therefore of interest as putative drug targets. One pathway in Ca(2+) homeostatic control in apicomplexans uses a Ca(2+)/H(+) exchanger (a member of the cation exchanger family, CAX). The P. falciparum CAX (PfCAX) has recently been characterised in asexual blood stage parasites. To determine the physiological importance of apicomplexan CAXs, tagging and knock-out strategies were undertaken in the genetically tractable T. gondii and P. berghei parasites. In addition, a yeast heterologous expression system was used to study the function of apicomplexan CAXs. Tagging of T. gondii and P. berghei CAXs (TgCAX and PbCAX) under control of their endogenous promoters could not demonstrate measureable expression of either CAX in tachyzoites and asexual blood stages, respectively. These results were consistent with the ability of parasites to tolerate knock-outs of the genes for TgCAX and PbCAX at these developmental stages. In contrast, PbCAX expression was detectable during sexual stages of development in female gametocytes/gametes, zygotes and ookinetes, where it was dispersed in membranous networks within the cytosol (with minimal mitochondrial localisation). Furthermore, genetically disrupted parasites failed to develop further from \"round\" form zygotes, suggesting that PbCAX is essential for ookinete development and differentiation. This impeded phenotype could be rescued by removal of extracellular Ca(2+). Therefore, PbCAX provides a mechanism for free living parasites to multiply within the ionic microenvironment of the mosquito midgut. Ca(2+) homeostasis mediated by PbCAX is critical and suggests plasmodial CAXs may be targeted in approaches designed to block parasite transmission.", "link"=>"http://www.mendeley.com/research/plasmodium-berghei-ca2h-exchanger-pbcax-essential-tolerance-environmental-ca2-during-sexual-developm", "reader_count"=>34, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>12, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>3, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>12, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>2, "Student > Master"=>4, "Other"=>1, "Student > Bachelor"=>3, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>24, "Medicine and Dentistry"=>3, "Philosophy"=>1, "Psychology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Psychology"=>{"Psychology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>24}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}, "Philosophy"=>{"Philosophy"=>1}}, "reader_count_by_country"=>{"Netherlands"=>1, "United Kingdom"=>2}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/974889"], "description"=>"<p>High resolution deconvolution microscopy images of a live female gametocyte shortly after activation and a female gamete/zygote and an ookinete 24 h post activation expressing PbCAX-GFP and co-stained with MitoTracker Red CMXRos (10 ng/ml) and Hoechst 33342. Scale bar: 5 µm.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>642797, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g004", "stats"=>{"downloads"=>1, "page_views"=>36, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PbCAX_GFP_localisation_/642797", "title"=>"PbCAX-GFP localisation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:20:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/974888"], "description"=>"<p>ΔpH-dependent uptake of 10 µM <sup>45</sup>Ca<sup>2+</sup> into vacuolar-enriched membrane vesicles isolated from K665 (<i>pmc1 vcx1</i>) yeast transformed with empty vector (piHGpd) (A), <i>scrcax1</i> (B), <i>pfcax</i> (C), and <i>spfcax</i> (D), measured over a 22 min time course. Transport measurements were determined in the presence of 0.1 mM NaN<sub>3</sub>, 10 mM KCl, 1 mM ATP, 1 mM MgSO<sub>4</sub> and 0.2 mM orthovanadate (a Ca<sup>2+</sup>-ATPase inhibitor). <sup>45</sup>Ca<sup>2+</sup> uptake in the absence (open symbols) or presence (closed symbols) of 5 µM of the protonophore CCCP is shown. The Ca<sup>2+</sup> ionophore A23187 was added at 12 min at a concentration of 5 µM to dissipate any vesicle-loaded <sup>45</sup>Ca<sup>2+</sup>, as indicated by arrows. Points represent the mean ± SEM of 4–5 independent experiments (where not shown errors lie within the symbols).</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>642796, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g003", "stats"=>{"downloads"=>1, "page_views"=>31, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ca_2_H_exchange_activity_of_PfCAX_/642796", "title"=>"Ca<sup>2+</sup>/H<sup>+</sup> exchange activity of PfCAX.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:19:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/974905"], "description"=>"<p>(A) Plaque assays were performed by incubating host cells with ΔKU80 and ΔCAX parasite strains for 7 days. (B) For the intracellular growth assay, ΔKU80 (open bars) and ΔCAX (closed bars) parasites were grown in host cells for 24 h and then the number of parasites per vacuole was determined. Bars represent the mean ± SEM of 3 independent experiments (each with 2 replicates). (C) For the ionophore-induced egress assay, parasites were cultured for 30 h before treatment with DMSO (open bars) or Ca<sup>2+</sup>-ionophore A23187 (3 µM; closed bars) for 5 min. The results are expressed as a percentage of ruptured vacuoles. Bars represent the mean ± SEM of at least 2 independent experiments.</p>", "links"=>[], "tags"=>["tgcax"], "article_id"=>642810, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g009", "stats"=>{"downloads"=>3, "page_views"=>49, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Essentiality_of_TgCAX_in_T_gondii_tachyzoites_/642810", "title"=>"Essentiality of TgCAX in <i>T. gondii</i> tachyzoites.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:22:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/974883"], "description"=>"<p>Amino acid sequence alignment of PfCAX with PbCAX. The Clustal W program was used to generate the alignment. The residues highlighted by a bold black line above correspond to transmembrane segment predictions determined with the TMHMM program (<a href=\"http://www.cbs.dtu.dk/services/TMHMM/\" target=\"_blank\">http://www.cbs.dtu.dk/services/TMHMM/</a>). The residues highlighted by a bold green line below correspond to the conserved CAX regions, c-1 and c-2. Green shading denotes residues shown to be essential for Ca<sup>2+</sup> transport in AtCAX1 and OsCAX1a <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003191#ppat.1003191-Kamiya1\" target=\"_blank\">[15]</a>, <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003191#ppat.1003191-Shigaki3\" target=\"_blank\">[16]</a>. Yellow shading denotes the putative mitochondrial targeting motif <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003191#ppat.1003191-Rotmann1\" target=\"_blank\">[7]</a>. Grey shading denotes cleaved sequences for mitochondrially imported proteins predicted by MitoProt II – v1.101 (<a href=\"http://ihg.gsf.de/ihg/mitoprot.html\" target=\"_blank\">http://ihg.gsf.de/ihg/mitoprot.html</a>). Red shading denotes phospo-acceptor sites (GeneDB and <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003191#ppat.1003191-Treeck1\" target=\"_blank\">[17]</a>). CAX sequences are from (accession no.): Pf, <i>Plasmodium falciparum</i> (XP_966025.1) and Pb, <i>Plasmodium berghei</i> (XP_678577.1). <i>Red letters</i>, identical or conserved residues in all sequences; <i>green letters</i>, conserved substitutions; <i>blue letters</i>, semi-conserved substitutions.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>642791, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sequence_alignments_/642791", "title"=>"Sequence alignments.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:19:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/974897"], "description"=>"<p>Line graph illustrating ookinete conversion, measured at 24 h post-gametocyte activation, in wild-type (WT, open circles: WT GFP, open triangles) and <i>Δpbcax cl9</i> (closed circles) and <i>cl5 gfp</i> (closed triangles) parasites in the presence of 10 mM EGTA added at 0, 0.5, 2 and 3 h post-gametocyte activation. The conversion rate is the percentage of P28-positive parasites that had successfully differentiated into elongated ‘banana-shaped’ ookinetes. Data are expressed as the percentage of wild-type controls. Points represent the mean ± SEM (n = 3) except for WT, where the points represent the mean ± range (n = 2).</p>", "links"=>[], "tags"=>["parasite"], "article_id"=>642805, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g007", "stats"=>{"downloads"=>0, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_916_pbcax_parasite_rescue_with_EGTA_/642805", "title"=>"<i>Δpbcax</i> parasite rescue with EGTA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:20:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/974896"], "description"=>"<p>Immunofluorescence images of <i>in vitro</i> wild-type (WT) and <i>Δpbcax cl9 P. berghei</i> ookinete cultures, 8 and 24 h after gametocyte activation and in the presence of EGTA (10 mM; added directly prior to gametocyte activation). Parasites are immunostained for the female gamete/zygote/ookinete marker P28 (red) and co-stained with the nuclear marker Hoechst 33342 (blue). Development of elongated ookinetes was completely ablated in the <i>Δpbcax cl9</i> line, a phenotype which could be reversed by the removal of extracellular Ca<sup>2+</sup> using EGTA. The arrow indicates the diffuse DNA staining observed in <i>Δpbcax cl9</i> parasites 24 h after gametocyte activation. Scale bar: 5 µm.</p>", "links"=>[], "tags"=>["parasite", "ookinete"], "article_id"=>642804, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g006", "stats"=>{"downloads"=>1, "page_views"=>40, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_916_pbcax_parasite_ookinete_conversion_images_/642804", "title"=>"<i>Δpbcax</i> parasite ookinete conversion images.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:20:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/974893"], "description"=>"<p>(A) Bar graph illustrating exflagellation in wild-type (WT) and <i>Δpbcax cl9</i> parasites. Exflagellation is presented as the numbers of exflagellation centres in 8 fields of view (magnification, ×40). Bars represent the mean ± SEM of 3 independent experiments. (B) Bar graph illustrating ookinete conversion in wild-type (WT or WT GFP) and <i>Δpbcax cl9 and cl5 gfp</i> parasites. The conversion rate is the percentage of P28-positive parasites that had successfully differentiated into elongated ‘banana-shaped’ ookinetes. Bars represent the mean ± SEM of 3–4 independent experiments except for WT GFP, where the bar represents the mean ± range (n = 2). (C) Ookinete conversion after crossing <i>Δpbcax cl9</i> parasites with female-defective <i>nek2</i> and <i>nek4</i> mutants (<i>Δpbnek2/4</i>) and a male-defective <i>map2</i> mutant (<i>Δpbmap2</i>). Wild-type parasites (WT) were used as a control. The conversion rate is the percentage of P28-positive parasites that had successfully differentiated into elongated ‘banana-shaped’ ookinetes. Bars represent the mean ± SEM of 3 independent experiments. (D) Bar graph illustrating the mean ± SEM numbers of oocysts per midgut (20 analysed) of wild-type (WT or WT GFP) and either <i>Δpbcax cl9</i> or <i>cl5 gfp</i> infected mosquitoes in three independent experiments.</p>", "links"=>[], "tags"=>["parasite"], "article_id"=>642801, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g005", "stats"=>{"downloads"=>2, "page_views"=>33, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_916_pbcax_parasite_phenotype_/642801", "title"=>"<i>Δpbcax</i> parasite phenotype.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:20:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/974915", "https://ndownloader.figshare.com/files/974916", "https://ndownloader.figshare.com/files/974917", "https://ndownloader.figshare.com/files/974918", "https://ndownloader.figshare.com/files/974919", "https://ndownloader.figshare.com/files/974920", "https://ndownloader.figshare.com/files/974921", "https://ndownloader.figshare.com/files/974922", "https://ndownloader.figshare.com/files/974923", "https://ndownloader.figshare.com/files/974924", "https://ndownloader.figshare.com/files/974925"], "description"=>"<div><p>Ca<sup>2+</sup> contributes to a myriad of important cellular processes in all organisms, including the apicomplexans, <i>Plasmodium</i> and <i>Toxoplasma</i>. Due to its varied and essential roles, free Ca<sup>2+</sup> is tightly regulated by complex mechanisms. These mechanisms are therefore of interest as putative drug targets. One pathway in Ca<sup>2+</sup> homeostatic control in apicomplexans uses a Ca<sup>2+</sup>/H<sup>+</sup> exchanger (a member of the cation exchanger family, CAX). The <i>P. falciparum</i> CAX (PfCAX) has recently been characterised in asexual blood stage parasites. To determine the physiological importance of apicomplexan CAXs, tagging and knock-out strategies were undertaken in the genetically tractable <i>T. gondii</i> and <i>P. berghei</i> parasites. In addition, a yeast heterologous expression system was used to study the function of apicomplexan CAXs. Tagging of <i>T. gondii</i> and <i>P. berghei</i> CAXs (TgCAX and PbCAX) under control of their endogenous promoters could not demonstrate measureable expression of either CAX in tachyzoites and asexual blood stages, respectively. These results were consistent with the ability of parasites to tolerate knock-outs of the genes for TgCAX and PbCAX at these developmental stages. In contrast, PbCAX expression was detectable during sexual stages of development in female gametocytes/gametes, zygotes and ookinetes, where it was dispersed in membranous networks within the cytosol (with minimal mitochondrial localisation). Furthermore, genetically disrupted parasites failed to develop further from “round” form zygotes, suggesting that PbCAX is essential for ookinete development and differentiation. This impeded phenotype could be rescued by removal of extracellular Ca<sup>2+</sup>. Therefore, PbCAX provides a mechanism for free living parasites to multiply within the ionic microenvironment of the mosquito midgut. Ca<sup>2+</sup> homeostasis mediated by PbCAX is critical and suggests plasmodial CAXs may be targeted in approaches designed to block parasite transmission.</p> </div>", "links"=>[], "tags"=>["development"], "article_id"=>642818, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1003191.s001", "https://dx.doi.org/10.1371/journal.ppat.1003191.s002", "https://dx.doi.org/10.1371/journal.ppat.1003191.s003", "https://dx.doi.org/10.1371/journal.ppat.1003191.s004", "https://dx.doi.org/10.1371/journal.ppat.1003191.s005", "https://dx.doi.org/10.1371/journal.ppat.1003191.s006", "https://dx.doi.org/10.1371/journal.ppat.1003191.s007", "https://dx.doi.org/10.1371/journal.ppat.1003191.s008", "https://dx.doi.org/10.1371/journal.ppat.1003191.s009", "https://dx.doi.org/10.1371/journal.ppat.1003191.s010", "https://dx.doi.org/10.1371/journal.ppat.1003191.s011"], "stats"=>{"downloads"=>10, "page_views"=>31, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_Plasmodium_berghei_Ca_2_H_Exchanger_PbCAX_Is_Essential_for_Tolerance_to_Environmental_Ca_2_during_Sexual_Development__/642818", "title"=>"The <em>Plasmodium berghei</em> Ca<sup>2+</sup>/H<sup>+</sup> Exchanger, PbCAX, Is Essential for Tolerance to Environmental Ca<sup>2+</sup> during Sexual Development", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-03-01 15:26:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/974903"], "description"=>"<p>(A) Immunofluorescence assay of intracellular <i>Toxoplasma</i> tachyzoites transiently transfected with <i>TgCAX-Ty</i> expressed under the control of the tubulin promotor (upper panels) and when expressed stably (lower panels). Parasites are immunostained with the surface marker GAP45 (red). (B) TgCAX transiently transfected tachyzoites detected by anti-TgCAX antibodies and co-stained with anti-Ty antibodies and the nuclear marker DAPI (upper panels) or with GAP45 antibodies (lower panels). (C) TgCAX transiently expressed in tachyzoites co-localised with 2 mitochondrial markers, anti-HSP70 antibodies <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003191#ppat.1003191-Pino1\" target=\"_blank\">[26]</a> (upper panels) or transiently co-transfected SPTPSOD2GFP (SP: signal peptide, TP: transit peptide, <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003191#ppat.1003191-Pino2\" target=\"_blank\">[27]</a>) (lower panels). Scale bars: 2 µm.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology"], "article_id"=>642809, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g008", "stats"=>{"downloads"=>0, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TgCAX_localisation_/642809", "title"=>"TgCAX localisation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:21:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/974886"], "description"=>"<p>(A) RT-PCR analysis of <i>pfcax</i> and <i>spfcax</i> expression in yeast compared with yeast transformed with the empty vector control. (B) Saturated liquid cultures of K665 (<i>pmc1 vcx1</i>) yeast transformed with <i>pfcax</i> in piHGpd, N-terminally truncated s<i>pfcax</i> in piHGpd, <i>scrcax1</i> in piHGpd and empty vector alone were serially diluted to the cell densities as indicated, then spotted onto selection medium lacking histidine (SD –His) and YPD medium containing 50 mM CaCl<sub>2</sub>. Yeast growth at 30°C is shown after 4 days. A representative experiment is shown. (C) K665 yeast transformed with the various plasmids described in (B) were diluted to a cell density of 0.5 <i>A</i><sub>600</sub> nm and inoculated into YPD medium containing concentrations of CaCl<sub>2</sub> from 25 to 150 mM. Yeast cell density was determined by absorbance measurements at 600 nm following growth shaking at 30°C for 16 h. Bars represent the mean ± SEM of 4 independent experiments (each with 8–12 replicates). Vector only, open bars; sCrCAX, closed bars; PfCAX, horizontally lined bars; sPfCAX, diagonally lined bars.</p>", "links"=>[], "tags"=>["yeast", "mediated"], "article_id"=>642794, "categories"=>["Microbiology", "Infectious Diseases"], "users"=>["David S. Guttery", "Jon K. Pittman", "Karine Frénal", "Benoit Poulin", "Leon R. McFarlane", "Ksenija Slavic", "Sally P. Wheatley", "Dominique Soldati-Favre", "Sanjeev Krishna", "Rita Tewari", "Henry M. Staines"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003191.g002", "stats"=>{"downloads"=>2, "page_views"=>33, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ca_2_tolerance_of_yeast_mediated_by_PfCAX_/642794", "title"=>"Ca<sup>2+</sup> tolerance of yeast mediated by PfCAX.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-03-01 15:19:40"}

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Relative Metric

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