Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist Following Loss of the Relict Plastid or Golgi Body Disruption
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{"title"=>"Vesicles bearing toxoplasma apicoplast membrane proteins persist following loss of the relict plastid or golgi body disruption", "type"=>"journal", "authors"=>[{"first_name"=>"Anne", "last_name"=>"Bouchut", "scopus_author_id"=>"6506036217"}, {"first_name"=>"Jennifer A.", "last_name"=>"Geiger", "scopus_author_id"=>"35298942200"}, {"first_name"=>"Amy E.", "last_name"=>"Derocher", "scopus_author_id"=>"7005032948"}, {"first_name"=>"Marilyn", "last_name"=>"Parsons", "scopus_author_id"=>"7202453240"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84954426985", "pui"=>"608758764", "doi"=>"10.1371/journal.pone.0112096", "issn"=>"19326203", "scopus"=>"2-s2.0-84954426985", "pmid"=>"25369183"}, "id"=>"29cd0369-3045-3c2c-ac4b-85d036746727", "abstract"=>"Toxoplasma gondii and malaria parasites contain a unique and essential relict plastid called the apicoplast. Most apicoplast proteins are encoded in the nucleus and are transported to the organelle via the endoplasmic reticulum (ER). Three trafficking routes have been proposed for apicoplast membrane proteins: (i) vesicular trafficking from the ER to the Golgi and then to the apicoplast, (ii) contiguity between the ER membrane and the apicoplast allowing direct flow of proteins, and (iii) vesicular transport directly from the ER to the apicoplast. Previously, we identified a set of membrane proteins of the T. gondii apicoplast which were also detected in large vesicles near the organelle. Data presented here show that the large vesicles bearing apicoplast membrane proteins are not the major carriers of luminal proteins. The vesicles continue to appear in parasites which have lost their plastid due to mis-segregation, indicating that the vesicles are not derived from the apicoplast. To test for a role of the Golgi body in vesicle formation, parasites were treated with brefeldin A or transiently transfected with a dominant-negative mutant of Sar1, a GTPase required for ER to Golgi trafficking. The immunofluorescence patterns showed little change. These findings were confirmed using stable transfectants, which expressed the toxic dominant-negative sar1 following Cre-loxP mediated promoter juxtaposition. Our data support the hypothesis that the large vesicles do not mediate the trafficking of luminal proteins to the apicoplast. The results further show that the large vesicles bearing apicoplast membrane proteins continue to be observed in the absence of Golgi and plastid function. These data raise the possibility that the apicoplast proteome is generated by two novel ER to plastid trafficking pathways, plus the small set of proteins encoded by the apicoplast genome.", "link"=>"http://www.mendeley.com/research/vesicles-bearing-toxoplasma-apicoplast-membrane-proteins-persist-following-loss-relict-plastid-golgi", "reader_count"=>16, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>5, "Student > Master"=>2, "Student > Bachelor"=>3, "Professor"=>1, "Student > Doctoral Student"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>5, "Student > Master"=>2, "Student > Bachelor"=>3, "Professor"=>1, "Student > Doctoral Student"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>7, "Agricultural and Biological Sciences"=>7, "Medicine and Dentistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>7}}, "group_count"=>0}

Scopus | Further Information

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  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934129/Figure_1.tif"], "description"=>"<p>For IFA analysis here and elsewhere unless indicated, proteins were detected by mAbs directed against epitope tags followed by fluorochrome-coupled secondary antibodies as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112096#s4\" target=\"_blank\">Methods</a>. In this case, the apicoplast membrane proteins were detected anti-HA mAb was followed by FITC-coupled secondary antibodies and <sup>S+T</sup>Red-V5 was detected by anti-V5 mAb followed by Texas Red-coupled antibodies to bypass the need for maturation of the HcRed chromophore. Here, as in other figures, the color coding for merged images is indicated by the text color above the merged images, while dashed lines mark the outline of the parasite. In this experiment, the parasites co-expressed <sup>S+T</sup>Red-V5 driven by the <i>ACP</i> promoter and epitope-tagged ApV proteins APT1-HA or ATrx1-HA. A) IFA showing the pattern seen in about 80% of parasites with ATrx1-HA in V<sup>ap</sup> (arrows) near the apicoplast. One set of anti-V5 images is scaled normally and the second is scaled to detect fainter signals (the staining at the apicoplast is then saturated). No evidence of localization of the luminal marker <sup>S+T</sup>Red-V5 with V<sup>ap</sup> was observed when scanning through the deconvolved planes. “H” marks a host cell nucleus. B) In the approximately 20% parasites with evident V<sup>ap</sup>, occasional regions staining for membrane-associated proteins (V<sup>ap</sup>, arrows) also showed a weak signal for the luminal marker <sup>S+T</sup>Red-V5. Bar, 2 µm. C) Individual parasites with V<sup>ap</sup> as detected by the presence of ATrx1-HA were randomly chosen for quantitation of ATrx1-HA and <sup>S+T</sup>Red-V5 signals. The average fluorescence corresponding to each protein in a 90 pixel area covering either the apicoplast (AP), vesicles (V<sup>ap</sup>) or adjacent regions (control) was determined and plotted for each individual parasite (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112096#s4\" target=\"_blank\">Methods</a>). The mean florescence signal seen in the parasite population is marked for each region analyzed (black lines). The raw fluorescence intensities for the adjacent regions averaged 12243 fluorescence units for ATrx1-HA and 5785 for <sup>S+T</sup>Red-V5, very close to the average background of 12267 (anti-HA, green line) and 6577 (anti-V5, red line) for these channels in untransfected RH parasites on the same slide.</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324757, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_V_ap_are_not_major_vehicles_for_luminal_protein_trafficking_to_the_apicoplast_/1324757", "title"=>"V<sup>ap</sup> are not major vehicles for luminal protein trafficking to the apicoplast.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934137/Figure_2.tif"], "description"=>"<p><i>T. gondii</i> expressing the indicated tagged apicoplast proteins were transiently transfected with a plasmid encoding <sup>S+T</sup>YFP-ROP1 (chimera, endogenous fluorescence) to induce plastid mis-segregation. After 40 hours to allow for apicoplast loss through several cell divisions, the samples were subjected to IFA. Vacuoles with one or more parasites expressing the chimeric protein were analyzed. Individual cells and vacuoles are outlined with solid lines and dashed lines respectively. The markers are indicated above each panel. DIC, differential interference contrast, H indicates host cell nucleus. A) Loss of luminal marker in parasites expressing the “poison” chimera. <sup>S+T</sup>Red-V5 was detected with both anti-V5 mAb (followed by secondary antibody coupled to Dylight 649; panels labeled <sup>S+T</sup>Red-V5), and through intrinsic fluorescence (panels here and in B, C labeled <sup>S+T</sup>Red). The lower panels show enhanced scaling of <sup>S+T</sup>Red-V5 detected with anti-V5 to highlight faint ER-like staining. Bar = 5 µM. B) Continued formation of V<sup>ap</sup> bearing FtsH1. FtsH1/<sup>S+T</sup>Red parasites were transiently transfected with the chimeric construct (detected by endogenous fluorescence) and FtsH1 was detected with anti-V5 mAb (followed by secondary antibody coupled to DyLight 649). Background of the <sup>S+T</sup>Red images in panels B and C were adjusted to correct for crossover fluorescence from the DyLight 649 fluorophore. Arrows indicate V<sup>ap</sup>-like staining in cells lacking an apicoplast. Vacuoles bearing transfected parasites (upper left) and untransfected parasites (lower right) are shown. Bar = 5 µM. C) Continued formation of V<sup>ap</sup> bearing ATrx1. ATrx1/<sup>S+T</sup>Red expressing cells were transiently transfected with <sup>S+T</sup>ROP1-YFP, which was detected by endogenous fluorescence. ATrx1 was detected with anti-HA mAb coupled to DyLight 649. Arrows indicate V<sup>ap</sup>-like staining in cells lacking an apicoplast. Vacuoles bearing transfected parasites (upper left) and untransfected parasites (lower right) are shown. Parasites in the lower vacuole are in stage 1 of the organelle division cycle and therefore have few V<sup>ap</sup>. Bar = 5 µM. D) Quantitation of V<sup>ap</sup> in apicoplast-deficient parasites. ATrx1-4HA/<sup>S+T</sup>Red and FtsH1-V5<sup>233</sup>-HA/<sup>S+T</sup>Red expressing cell lines were transiently transfected with the chimeric <sup>S+T</sup>YFP-ROP1 construct and vacuoles were scored for the presence or absence of YFP in at least one parasite (indicating expression of the chimeric protein in the original invading parasite). Individual parasites within each vacuole were then scored for the presence or absence of the luminal protein <sup>S+T</sup>Red (detected by endogenous fluorescence) and the apicoplast membrane protein (detected by anti-HA or anti-V5 mAbs followed by anti-mouse IgG coupled to DyLight 649). The bar graph plots the percentage of cells bearing each marker protein in vacuoles derived from transfected (chimera<sup>+</sup>) and untransfected (chimera<sup>−</sup>) parasites. In the ATrx1 sample, 96 chimera<sup>+</sup> and 128 chimera<sup>−</sup> cells were counted; in the FtsH1 sample, 27 chimera<sup>+</sup> and 48 chimera<sup>−</sup> cells were counted. These results are representative of three independent experiments.</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324759, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_V_ap_persist_in_parasites_with_plastid_loss_/1324759", "title"=>"V<sup>ap</sup> persist in parasites with plastid loss.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934139/Figure_3.tif"], "description"=>"<p>A) Parasites were incubated with or without 1 µg/ml BFA for 1 hour at 37°C. <i>T. gondii</i> co-expressing the Golgi matrix marker GRASP55-YFP and FtsH1 internally tagged with V5 epitopes were stained with anti-V5 mAb followed by secondary antibody coupled to DyLight 649 and Texas Red streptavidin (which detects a naturally biotinylated protein in the apicoplast lumen, AP lumen). Bar, 2 µM. B) Parasites expressing the Golgi membrane protein NST1-HA (detected with anti-HA mAb coupled to Alexa 594) served as the control, demonstrating the effectiveness of BFA. Bar, 2 µM. C) Quantitative analysis of signal overlap between internally tagged FtsH1 and GRASP55 in the presence or absence of BFA. More than 50 parasites were analyzed for each condition (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112096#s4\" target=\"_blank\">Methods</a>).</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324761, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Overlap_between_FtsH1_and_GRASP55_does_not_reflect_FtsH1_protein_in_the_Golgi_body_/1324761", "title"=>"Overlap between FtsH1 and GRASP55 does not reflect FtsH1 protein in the Golgi body.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934140/Figure_4.tif"], "description"=>"<p>Fibroblast monolayers infected with <i>T. gondii</i> expressing FtsH1 tagged internally with V5 epitopes and a cytosolic GFP (∼10<sup>8</sup>) were pre-incubated with or without BFA (1 µg/ml) for the indicated times prior to being labeled with <sup>35</sup>S-methionine/cysteine for 30 minutes. Samples were immunoprecipitated with anti-V5 mAb, anti-GFP, and anti-MIC5 before being separated on 7.5% (FtsH1) or 8–16% (GFP and MIC5) SDS-PAGE gels and transferred to nitrocellulose. The left panel shows phosphorimaging, the right panel shows the same lanes detected by Western blot. The four major forms of FtsH1 are marked according to their apparent molecular mass on SDS-PAGE: full-length (F-170), N-terminally processed (NP-154), C-terminally processed (CP-140) or dual processed (NPCP-115). In a 30 min labeling, the first two forms predominate <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112096#pone.0112096-Karnataki1\" target=\"_blank\">[21]</a>. The precursor (p) and mature (m) forms of MIC5 <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112096#pone.0112096-Brydges1\" target=\"_blank\">[75]</a> are marked.</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324762, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Protein_synthesis_during_BFA_treatment_assessed_by_biosynthetic_labeling_of_FtsH1_MIC5_and_cytosolic_GFP_/1324762", "title"=>"Protein synthesis during BFA treatment assessed by biosynthetic labeling of FtsH1, MIC5 and cytosolic GFP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934143/Figure_5.tif"], "description"=>"<p>A) IFA analysis of <i>T. gondii</i> grown with or without 1 µg/ml BFA for 1.5 hours. The parasites expressed the luminal apicoplast marker <sup>S+T</sup>Red along with FtsH1internally tagged with V5 epitopes or ATrx1-HA, which were detected with α-V5 followed by anti-mouse IgG (FITC) or anti-HA mAb directly coupled to FITC. Arrows indicate V<sup>ap</sup>-like structures in control and BFA-treated parasites. Bar, 2 µM. B) IFA of parasites expressing the Golgi membrane protein NST1-HA (detected with anti-HA mAb coupled to FITC), treated in parallel. Bar, 2 µM. C) Quantitation. The percentage of vacuoles with parasites bearing V<sup>ap</sup> in the presence (+) or absence (−) of BFA is depicted. Three replicates are shown for FtsH1 (circle) and one for ATrx1 (triangle). More than 125 vacuoles were analyzed for each point. At the times chosen, the proportion of parasites with apicoplasts at stage 2 (elongated oval), stage 3 (elongated bar), and stage 4 (V-shaped bar) were: FtsH1 analysis: control, 82.7%; BFA, 72.2%; ATrx1 analysis: control, 81.6%; BFA, 74.5%.</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324765, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_V_ap_persist_in_the_presence_of_the_Golgi_disruptor_BFA_/1324765", "title"=>"V<sup>ap</sup> persist in the presence of the Golgi disruptor BFA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934144/Figure_6.tif"], "description"=>"<p>Intracellular <i>T. gondii</i> expressing FtsH1 internally tagged withV5 epitopes were metabolically labeled for 30 min and then chased for various times in complete medium. BFA-treated samples included the drug throughout the pulse-chase. FtsH1 was then immunoprecipitated and subjected to SDS-PAGE followed by phosphorimaging (<sup>35</sup>S panel). The blot was subsequently probed with anti-V5 mAb (Western panel). The four major forms of FtsH1 are marked according to their apparent molecular mass on SDS-PAGE: full-length (F-170), N-terminally processed (NP-154), C-terminally processed (CP-140) or dual processed (NPCP-115).</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324766, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Effect_of_BFA_on_FtsH1_processing_/1324766", "title"=>"Effect of BFA on FtsH1 processing.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934146/Figure_7.tif"], "description"=>"<p>A) <i>T. gondii</i> expressing <sup>S+T</sup>Red plus ATrx1-HA or FtsH1 internally tagged with V5 epitopes were transiently transfected with either wt SAR1-GFP or sar1(H74L)-GFP and analyzed by IFA for the localization of the two ApV protein. Epitope tagged proteins were detected by mAbs reactive with the epitope tags followed by secondary antibodies coupled to DyLight 649. The fluorescent proteins were detected by endogenous fluorescence. Arrows point to V<sup>ap</sup>-like structures. Bar, 2 µM. B) Overexpression of sar1(H74L) abrogates localization of NST1. SAR1 and sar1(H74L) constructs were transiently transfected into <i>T. gondii</i> expressing NST1-HA and the samples analyzed as above. Note the reticular staining of NST1 following expression of the dominant negative protein. C) V<sup>ap</sup> are still present after induction of sar1(H74L) in stable transfectants. As described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112096#s4\" target=\"_blank\">Methods</a>, parasites were stably transfected with constructs bearing sar1(H74L)-YFP or the wt SAR1-YFP that was separated from a promoter by RFP flanked by <i>loxP</i> sequences. Addition of rapamycin led to excision of the RFP sequence that separated and expression of the test proteins. After 11 hours YFP<sup>+</sup> parasites were scored for the presence or absence of V<sup>ap</sup> using mAb 11G8 which detects ATrx1 (followed by secondary antibody coupled to DyLight 350).</p>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324768, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/_Expression_of_dominant_negative_sar1_does_not_eliminate_V_ap_/1324768", "title"=>"Expression of dominant negative <i>sar1</i> does not eliminate V<sup>ap</sup>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-04 22:04:10"}
  • {"files"=>["https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934153/Figure_S1.tif", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934154/Figure_S2.tif", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934155/Figure_S3.tif", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934156/Figure_S4.tif", "https://s3-eu-west-1.amazonaws.com/pstorage-plos-3567654/1934157/Figure_S5.tif"], "description"=>"<div><p><i>Toxoplasma gondii</i> and malaria parasites contain a unique and essential relict plastid called the apicoplast. Most apicoplast proteins are encoded in the nucleus and are transported to the organelle <i>via</i> the endoplasmic reticulum (ER). Three trafficking routes have been proposed for apicoplast membrane proteins: (i) vesicular trafficking from the ER to the Golgi and then to the apicoplast, (ii) contiguity between the ER membrane and the apicoplast allowing direct flow of proteins, and (iii) vesicular transport directly from the ER to the apicoplast. Previously, we identified a set of membrane proteins of the <i>T. gondii</i> apicoplast which were also detected in large vesicles near the organelle. Data presented here show that the large vesicles bearing apicoplast membrane proteins are not the major carriers of luminal proteins. The vesicles continue to appear in parasites which have lost their plastid due to mis-segregation, indicating that the vesicles are not derived from the apicoplast. To test for a role of the Golgi body in vesicle formation, parasites were treated with brefeldin A or transiently transfected with a dominant-negative mutant of Sar1, a GTPase required for ER to Golgi trafficking. The immunofluorescence patterns showed little change. These findings were confirmed using stable transfectants, which expressed the toxic dominant-negative sar1 following Cre-<i>loxP</i> mediated promoter juxtaposition. Our data support the hypothesis that the large vesicles do not mediate the trafficking of luminal proteins to the apicoplast. The results further show that the large vesicles bearing apicoplast membrane proteins continue to be observed in the absence of Golgi and plastid function. These data raise the possibility that the apicoplast proteome is generated by two novel ER to plastid trafficking pathways, plus the small set of proteins encoded by the apicoplast genome.</p></div>", "links"=>[], "tags"=>["luminal proteins", "er", "Vesicles Bearing Toxoplasma Apicoplast Membrane Proteins Persist", "vesicle", "plastid trafficking pathways", "Golgi Body Disruption Toxoplasma gondii", "apicoplast membrane proteins"], "article_id"=>1324775, "categories"=>["Uncategorised"], "users"=>["Anne Bouchut", "Jennifer A. Geiger", "Amy E. DeRocher", "Marilyn Parsons"], "doi"=>["http://dx.doi.org/10.1371/journal.pone.0112096"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"http://figshare.com/articles/Vesicles_Bearing_Toxoplasma_Apicoplast_Membrane_Proteins_Persist_Following_Loss_of_the_Relict_Plastid_or_Golgi_Body_Disruption/1324775", "title"=>"Vesicles Bearing <i>Toxoplasma</i> Apicoplast Membrane Proteins Persist Following Loss of the Relict Plastid or Golgi Body Disruption", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-11-04 22:04:10"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[291]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[286]}, {"subject_area"=>"/Biology and life sciences/Organisms", "average_usage"=>[310]}, {"subject_area"=>"/Biology and life sciences/Plant science", "average_usage"=>[278, 438]}]}
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