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dc.contributor.authorHutson, Samuel L.
dc.contributor.authorMui, Ernest
dc.contributor.authorKinsley, Karen
dc.contributor.authorWitola, William H.
dc.contributor.authorBehnke, Michael S.
dc.contributor.authorEl Bissati, Kamal
dc.contributor.authorMuench, Stephen P.
dc.contributor.authorRohrman, Brittany
dc.contributor.authorLiu, Susan R.
dc.contributor.authorWollmann, Robert
dc.contributor.authorOgata, Yuko
dc.contributor.authorSarkeshik, Ali
dc.contributor.authorYates III, John R.
dc.contributor.authorMcLeod, Rima
dc.date.accessioned2019-04-17T20:56:35Z
dc.date.available2019-04-17T20:56:35Z
dc.date.issued2010-11
dc.identifier.citationHutson, Samuel L., Ernest Mui, Karen Kinsley, William H. Witola, Michael S. Behnke, Kamal El Bissati, Stephen P. Muench, et al. “T. Gondii RP Promoters & Knockdown Reveal Molecular Pathways Associated with Proliferation and Cell-Cycle Arrest.” Edited by Gordon Langsley. PLoS ONE 5, no. 11 (November 22, 2010): e14057. doi:10.1371/journal.pone.0014057.en_US
dc.identifier.issn1932-6203
dc.identifier.urihttps://scholarworks.montana.edu/xmlui/handle/1/15439
dc.description.abstractMolecular pathways regulating rapid proliferation and persistence are fundamental for pathogens but are not elucidated fully in Toxoplasma gondii. Promoters of T. gondii ribosomal proteins (RPs) were analyzed by EMSAs and ChIP. One RP promoter domain, known to bind an Apetela 2, bound to nuclear extract proteins. Promoter domains appeared to associate with histone acetyl transferases. To study effects of a RP gene's regulation in T. gondii, mutant parasites (Δrps13) were engineered with integration of tetracycline repressor (TetR) response elements in a critical location in the rps13 promoter and transfection of a yellow fluorescent-tetracycline repressor (YFP-TetR). This permitted conditional knockdown of rps13 expression in a tightly regulated manner. Δrps13 parasites were studied in the presence (+ATc) or absence of anhydrotetracycline (-ATc) in culture. -ATc, transcription of the rps13 gene and expression of RPS13 protein were markedly diminished, with concomitant cessation of parasite replication. Study of Δrps13 expressing Myc-tagged RPL22, -ATc, showed RPL22 diminished but at a slower rate. Quantitation of RNA showed diminution of 18S RNA. Depletion of RPS13 caused arrest of parasites in the G1 cell cycle phase, thereby stopping parasite proliferation. Transcriptional differences ±ATc implicate molecules likely to function in regulation of these processes. In vitro, -ATc, Δrps13 persists for months and the proliferation phenotype can be rescued with ATc. In vivo, however, Δrps13 could only be rescued when ATc was given simultaneously and not at any time after 1 week, even when L-NAME and ATc were administered. Immunization with Δrps13 parasites protects mice completely against subsequent challenge with wildtype clonal Type 1 parasites, and robustly protects mice against wildtype clonal Type 2 parasites. Our results demonstrate that G1 arrest by ribosomal protein depletion is associated with persistence of T. gondii in a model system in vitro and immunization with Δrps13 protects mice against subsequent challenge with wildtype parasites.en_US
dc.description.sponsorshipFin Charity Trust; Rooney Aldens; Peter Mann and Dominique Cornwell Family Trust; Morel, Jackson, Orlinsky, Goldberg, Taub, Kapnick, Schilling, and Kiewit families; Intervet/Schering Plough; Toxoplasmosis Research Institute; Research to Prevent Blindness Foundation; Howard Hughes Foundation Undergraduate Summer Research Program in Neurosciences; National Institutes of Health National Institute of Allergy and Infectious Diseases R01AI043228, U01 AI082180-01, U01 AI 77887, R01 AI071319-01, NIH P41RR0118232en_US
dc.language.isoenen_US
dc.rightsCC BY: This license lets you distribute, remix, tweak, and build upon this work, even commercially, as long as you credit the original creator for this work. This is the most accommodating of licenses offered. Recommended for maximum dissemination and use of licensed materials.en_US
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/legalcodeen_US
dc.titleT. gondii RP promoters & knockdown reveal molecular pathways associated with proliferation and cell-cycle arresten_US
dc.typeArticleen_US
mus.citation.issue11en_US
mus.citation.journaltitlePLoS Oneen_US
mus.citation.volume5en_US
mus.identifier.categoryLife Sciences & Earth Sciencesen_US
mus.identifier.doi10.1371/journal.pone.0014057en_US
mus.relation.collegeCollege of Letters & Scienceen_US
mus.relation.departmentMicrobiology & Immunology.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.relation.researchgroupMT INBRE Bioinformatics and Biostatistics Core.en_US
mus.data.thumbpage6en_US


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CC BY: This license lets you distribute, remix, tweak, and build upon this work, even commercially, as long as you credit the original creator for this work. This is the most accommodating of licenses offered. Recommended for maximum dissemination and use of licensed materials.
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