title: Garcinol Inhibits GCN5-Mediated Lysine Acetyltransferase Activity and Prevents Replication of the Parasite Toxoplasma gondii.
abstract: Lysine acetylation is a critical posttranslational modification that influences protein activity, stability, and binding properties. The acetylation of histone proteins in particular is a well-characterized feature of gene expression regulation. In the protozoan parasiteToxoplasma gondii, a number of lysine acetyltransferases (KATs) contribute to gene expression and are essential for parasite viability. The natural product garcinol was recently reported to inhibit enzymatic activities of GCN5 and p300 family KATs in other species. Here we show that garcinol inhibits TgGCN5b, the only nuclear GCN5 family KAT known to be required forToxoplasmatachyzoite replication. Treatment of tachyzoites with garcinol led to a reduction of global lysine acetylation, particularly on histone H3 and TgGCN5b itself. We also performed transcriptome sequencing (RNA-seq), which revealed increasing aberrant gene expression coincident with increasing concentrations of garcinol. The majority of the genes that were most significantly affected by garcinol were also associated with TgGCN5b in a previously reported chromatin immunoprecipitation assay with microarray technology (ChIP-chip) analysis. The dysregulated gene expression induced by garcinol significantly inhibitsToxoplasmatachyzoite replication, and the concentrations used exhibit no overt toxicity on human host cells. Garcinol also inhibitsPlasmodium falciparumasexual replication with a 50% inhibitory concentration (IC50) similar to that forToxoplasma Together, these data support that pharmacological inhibition of TgGCN5b leads to a catastrophic failure in gene expression control that prevents parasite replication.
authors.affiliation: Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, Indiana, USA.
authors.forename: Victoria
authors.initials: V
authors.lastname: Jeffers
authors.affiliation: Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana, USA.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Gao
authors.affiliation: Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, USA.
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Checkley
authors.affiliation: Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, Indiana, USA.
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Ferdig
authors.affiliation: Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, Indiana, USA Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA wjsulliv@iu.edu.
authors.forename: William J
authors.initials: WJ
authors.lastname: Sullivan
mesh_terms: D000107:Acetylation; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects / Q000469:parasitology; D005347:Fibroblasts / Q000187:drug effects / Q000469:parasitology; D020869:Gene Expression Profiling; D051548:Histone Acetyltransferases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D008018:Life Cycle Stages / Q000187:drug effects / Q000235:genetics; D008239:Lysine / Q000378:metabolism; D046228:Microarray Analysis; D058977:Molecular Sequence Annotation; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA; D013729:Terpenes / Q000494:pharmacology*; D014122:Toxoplasma / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome; D000099224:p300-CBP-Associated Factor
chemical_list: D000981:Antiprotozoal Agents; D051548:Histone Acetyltransferases; D006657:Histones; D008239:Lysine; D015800:Protozoan Proteins; D013729:Terpenes; C402103:GCN5 protein, Toxoplasma gondii; D000099224:p300-CBP-Associated Factor; C054597:garcinol
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