The protozoan parasite Toxoplasma gondii encodes a gamut of phosphodiesterases during its lytic cycle in human cells
| Autor: | Özlem Günay-Esiyok, Nicolas Liem, Kim Chi Vo, Nishith Gupta |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
PDE
phosphodiesterase AC adenylate cyclase COS crossover sequence Biochemistry chemistry.chemical_compound 0302 clinical medicine Structural Biology QMEAN Quality Model Energy Analysis MOI multiplicity of infection smHA spaghetti monster-HA cAMP and cGMP signaling 3′IT 3′-insertional tagging 0303 health sciences Lytic cycle OCRE octamer repeat Phosphodiesterase Computer Science Applications Cell biology HFF human foreskin fibroblast HXGPRT hypoxanthine-xanthine-guanine phosphoribosyltransferase CRISPR clustered regularly interspaced short palindromic repeats 030220 oncology & carcinogenesis PM plasma membrane GC guanylate cyclase Biotechnology Research Article Bradyzoite lcsh:Biotechnology Biophysics Mutagenesis (molecular biology technique) Biology Apicomplexa 03 medical and health sciences Cyclic nucleotide lcsh:TP248.13-248.65 GMQE Global Model Quality Estimation Genetics 030304 developmental biology ComputingMethodologies_COMPUTERGRAPHICS FPKM fragments per kilobase of exon model per million Tachyzoite EES entero-epithelial stages Intracellular parasite sgRNA single guide RNA Toxoplasma gondii Subcellular localization biology.organism_classification chemistry IMC inner membrane complex MAEBL merozoite adhesive erythrocytic binding ligand PKG protein kinase G PKA protein kinase A |
| Zdroj: | Computational and Structural Biotechnology Journal Computational and Structural Biotechnology Journal, Vol 18, Iss, Pp 3861-3876 (2020) |
| ISSN: | 2001-0370 |
| Popis: | Graphical abstract Highlights • Toxoplasma genome harbors at least 18 phosphodiesterases encoded by distinct genes. • Most parasite PDEs lack regulatory modules and are quite divergent from their human orthologs. • Acutely-infectious tachyzoite stage of T. gondii expresses 11 PDEs with varied localizations. • PDE8 and PDE9 are closely-related dual-substrate specific proteins residing in the apical pole. • Homology modeling of PDE8 and PDE9 reveals a conserved 3D topology and substrate pocket. • PDE9 is dispensable in tachyzoites, signifying a functional redundancy with PDE8. Cyclic nucleotide signaling is pivotal to the asexual reproduction of Toxoplasma gondii, however little do we know about the phosphodiesterase enzymes in this widespread obligate intracellular parasite. Here, we identified 18 phosphodiesterases (TgPDE1-18) in the parasite genome, most of which form apicomplexan-specific clades and lack archetypal regulatory motifs often found in mammalian PDEs. Genomic epitope-tagging in the tachyzoite stage showed the expression of 11 phosphodiesterases with diverse subcellular distributions. Notably, TgPDE8 and TgPDE9 are located in the apical plasma membrane to regulate cAMP and cGMP signaling, as suggested by their dual-substrate catalysis and structure modeling. TgPDE9 expression can be ablated with no apparent loss of growth fitness in tachyzoites. Likewise, the redundancy in protein expression, subcellular localization and predicted substrate specificity of several other PDEs indicate significant plasticity and spatial control of cyclic nucleotide signaling during the lytic cycle. Our findings shall enable a rational dissection of signaling in tachyzoites by combinatorial mutagenesis. Moreover, the phylogenetic divergence of selected Toxoplasma PDEs from human counterparts can be exploited to develop parasite-specific inhibitors and therapeutics. |
| Databáze: | OpenAIRE |
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