3160 total record number
71 records this year

To narrow your search, use one or more of the following search menus below.

To search by keyword, you may search by type of cell/animal/assay/protein/research or publication.

An unusual and physiologically vital protein with guanylate cyclase and P-type ATPase like domain in a pathogenic protist:

Günay-Esiyok, ;Scheib, U;Noll, M;Gupta, N;
Product: Alpha Toxin from C. septicum, Liquid

Cyclic GMP is considered as one of the master regulators of diverse functions in eukaryotes; its architecture and functioning in protozoans remain poorly understood however. We characterized an unusual and extra-large guanylate cyclase (477-kDa) containing at least 4 putative P-type ATPase motifs and 21 transmembrane helices in a common parasitic protist, Toxoplasma gondii. This protein, termed as TgATPaseP-GC due to its anticipated multi-functionality, localizes in the plasma membrane at the apical pole, while the corresponding cGMP-dependent protein kinase (TgPKG) is distributed in cytomembranes. Both proteins are expressed constitutively during the entire lytic cycle of the parasite in human cells, which suggests a post-translational control of cGMP signaling. Homology modeling indicated an activation of guanylate cyclase by heterodimerization of its two cyclase domains. TgATPaseP-GC is refractory to genetic deletion, and its CRISPR/Cas9-mediated disruption aborts the lytic cycle. Likewise, Cre/loxP-regulated knockdown of the TgATPaseP-GC by 3-UTR excision inhibited the parasite growth due to impairments in motility-dependent egress and invasion events. Consistently, cGMP-specific phosphodiesterase inhibitors restored the gliding motility of the mutant. A genetic repression of TgPKG, or its pharmacological inhibition phenocopied the defects observed in the TgATPaseP-GC mutant. Our data show a vital function of cGMP signaling, which is inducted by an alveolate-specific guanylate cyclase coupled to P-type like ATPase, and transduced by a dedicated PKG in T. gondii. The presence of TgATPaseP-GC orthologs in many other alveolates with contrasting habitats implies a divergent functional repurposing of cGMP signaling in protozoans. The work also lays an avenue to systematically dissect the cascade and understand its evolution in a model protist.