The protozoan parasite Toxoplasma gondii encodes a gamut of phosphodiesterases during its lytic cycle in human cells.
Comput Struct Biotechnol J
; 18: 3861-3876, 2020.
Article
en En
| MEDLINE
| ID: mdl-33335684
ABSTRACT
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.
3'IT, 3'-insertional tagging; AC, adenylate cyclase; Apicomplexa; Bradyzoite; COS, crossover sequence; CRISPR, clustered regularly interspaced short palindromic repeats; EES, entero-epithelial stages; FPKM, fragments per kilobase of exon model per million; GC, guanylate cyclase; GMQE, Global Model Quality Estimation; HFF, human foreskin fibroblast; HXGPRT, hypoxanthine-xanthine-guanine phosphoribosyltransferase; IMC, inner membrane complex; Lytic cycle; MAEBL, merozoite adhesive erythrocytic binding ligand; MOI, multiplicity of infection; OCRE, octamer repeat; PDE, phosphodiesterase; PKA, protein kinase A; PKG, protein kinase G; PM, plasma membrane; QMEAN, Quality Model Energy Analysis; Tachyzoite; cAMP and cGMP signaling; sgRNA, single guide RNA; smHA, spaghetti monster-HA
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Comput Struct Biotechnol J
Año:
2020
Tipo del documento:
Article
País de afiliación:
Alemania
Pais de publicación:
HOLANDA
/
HOLLAND
/
NETHERLANDS
/
NL
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PAISES BAJOS
/
THE NETHERLANDS