| 2014 |
ROP5 complexes with the active kinases ROP18 and ROP17, as demonstrated by tandem affinity protein tagging and purification in T. gondii. Biochemical analyses indicate that ROP18 and ROP17 have evolved to target adjacent and essential threonine residues in switch region I of immunity-related GTPases (IRGs). The combined activities of ROP17 and ROP18 contribute to avoidance of IRG recruitment to the intracellular T. gondii-containing vacuole, protecting the parasite from clearance in interferon-activated macrophages. |
Tandem affinity purification (TAP tagging), Co-IP, biochemical kinase assay, IRG recruitment assay in interferon-activated macrophages |
Cell host & microbe |
High |
24832449
|
| 2014 |
ROP5 binds adjacent to the nucleotide-binding domain of an IRG, producing an allosteric change that alters the orientation of two threonine residues targeted by Toxoplasma kinases ROP17 and ROP18, thereby facilitating their phosphorylation by ROP17/ROP18. |
Structural and biochemical analyses of ROP5-IRG interaction, allosteric conformational analysis |
The Journal of biological chemistry |
High |
25118287
|
| 2019 |
ROP17 is required for enhanced tissue migration of parasitized monocytes and macrophages through collagen-rich tissues in a Rho-ROCK-dependent manner consistent with integrin-independent interstitial migration. ROP17-deficient parasites failed to upregulate monocyte tissue migration and exhibited an early dissemination delay in vivo. |
ROP17 knockout parasites, in vitro monocyte migration assays (endothelial and collagen matrix), in vivo mouse dissemination assay |
Nature microbiology |
High |
31332383
|
| 2019 |
ROP17 kinase activity is required for translocation of dense granule (GRA) effector proteins across the parasitophorous vacuolar membrane (PVM). Null or catalytically dead (kinase-dead) mutants of ROP17 are defective in GRA translocation. ROP17 resides on the host cytosol side of the PVM and carries out its translocation function from this location, as demonstrated by trans-complementation with ROP17 delivered by co-infecting tachyzoites. |
Whole-genome sequencing of translocation-defective mutants, null mutant and catalytically dead mutant analysis, trans-complementation assay, subcellular localization |
mSphere |
High |
31366709
|
| 2020 |
ROP17 was identified as one of five parasite proteins required for translocation of GRA effectors across the PVM, alongside MYR1, MYR2, MYR3, and ASP5. Co-immunoprecipitation of MYR1 confirmed its association within this translocation complex. |
Affinity purification of MYR1 (Co-IP), genetic knockouts, GRA16 translocation assay |
mSphere |
High |
32075880
|
| 2016 |
Deletion of ROP17 in type II T. gondii resulted in a >12-fold defect in virulence in mice; however, resistance to host IRGs was not affected in the Δrop17 strain. Combined Δrop17/Δrop5 and Δrop17/Δrop18 showed that ROP17 contributes to virulence via an IRG-independent mechanism. Deletion of ROP17 severely reduced tissue cyst burdens in chronic infection. |
Gene knockout (targeted deletion of 26 ROPK loci), mouse virulence assay, IRG recruitment assay, cyst burden quantification |
mBio |
High |
27165797
|
| 2016 |
Combined Δrop21/Δrop17 double knockout led to a 50% reduction in cyst burden in vivo in chronic infection, whereas single Δrop17 knockout had no effect on in vitro growth or bradyzoite differentiation. |
CRISPR/Cas9 gene deletion, mouse cyst burden assay, in vitro growth assay |
Cellular microbiology |
Medium |
27450947
|
| 2020 |
GRA60, a dense granule protein, associates with ROP18 (a member of the ROP5/ROP17/ROP18 virulence complex) and contributes to resistance against IRG recruitment to the PVM. Deletion of GRA60 in RHΔrop5 background showed IRG recruitment comparable to single knockouts, indicating GRA60 functions in parallel with or upstream of the ROP5/ROP17/ROP18 complex. |
Genetic knockout, Co-immunoprecipitation (GRA60 with ROP18), IRG recruitment assay (Irgb10, Irga6) |
Cellular microbiology |
Medium |
33040458
|
| 2016 |
Deletion of ROP17 in a type II T. gondii strain severely abrogated the antitumor immune response induced by uracil auxotrophic mutants, indicating ROP17 plays a role in activating host innate immune responses through parasitophorous vacuole membrane-associated functions. |
Targeted gene deletion, tumor immunotherapy mouse model, antitumor immunity assay |
PLoS genetics |
Medium |
27447180
|
| 2014 |
TgROP17 expressed in HEK 293T cells phosphorylates c-Jun, demonstrating serine/threonine kinase activity. ROP17 overexpression also inhibited camptothecin-induced apoptosis. |
Eukaryotic expression vector transfection, Western blotting for c-Jun phosphorylation, flow cytometry apoptosis assay |
Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi |
Low |
24822361
|
| 2016 |
ROP17 overexpression in J774A.1 macrophages inhibited IFN-γ-induced apoptosis via phosphorylation of c-Jun and upregulation of Bcl-3; this anti-apoptotic effect was dependent on c-Jun phosphorylation as shown by c-Jun shRNA knockdown. |
Plasmid transfection, flow cytometry, Western blotting for phospho-c-Jun, Bcl-3, cleaved Caspase-3, c-Jun shRNA knockdown |
Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi |
Low |
30141842
|
| 2021 |
ROP17 interacts with Bcl-2 as demonstrated by co-immunoprecipitation, and ROP17 overexpression promotes autophagy via the Bcl-2–Beclin 1 pathway in HEK 293T cells. |
Co-immunoprecipitation, MDC staining, transmission electron microscopy, fluorescence microscopy, Western blotting, immunohistochemistry in vivo |
Folia parasitologica |
Low |
34180401
|
| 2016 |
Deletion of ROP17 individually in a Chinese type I strain (T.gHB1) showed similar virulence attenuation as in the RH strain, but double deletions of GRA7 plus ROP17 in T.gHB1 did not show significant further attenuation compared to the ROP17 single knockout, suggesting ROP17 and GRA7 may not act independently in all strain backgrounds. |
CRISPR-Cas9 gene deletion, mouse virulence assay, double knockout epistasis analysis |
Parasitology research |
Medium |
27225000
|