RhopH3 phosphorylation at S804 is important for its localization to the rhoptries. (A to C) IFA was performed on schizont-stage 3D7 or R3_S804A parasites to stain RhopH3 along with RAP1 (A), Clag3.1 (B), and AMA1 (C). While RhopH3 staining was observed in rhoptries as indicated by punctae proximal to RAP1 (A) or colocalized with Clag3.1 (B) in 3D7 parasites, it was significantly diffused in parasite cytoplasm of mutant parasites. The localization of microneme protein AMA1 was unchanged in the mutant parasites (C).RhopH3 was localized as puncta adjacent to rhoptry bulb protein RAP1 as expected in 3D7 parasites (A). In sharp contrast, RhopH3 S804A mutant was diffused and did not exhibit exclusive punctae typical of rhoptries in a significant number of parasites (A). RAP1 staining was almost unaltered in these parasites (A) Clag3.1 exhibited expected punctate rhoptry localization which colocalized with RhopH3 in 3D7 parasites (B). In most R3_S804A parasites, Clag3.1 showed a similar pattern whereas RhopH3 was more diffuse in the cytoplasm and exhibited significant reduction in colocalization with Clag3.1 (B). The localization of AMA1 to micronemes, which is reflected by punctate apical staining, was also unchanged upon mutation of S804A (C) whereas RhopH3 was significantly mislocalized to the parasite cytoplasm (C).
Ekka R, Gupta A, Bhatnagar S, Malhotra P, Sharma P. Phosphorylation of Rhoptry Protein RhopH3 Is Critical for Host Cell Invasion by the Malaria Parasite. mBio. 2020 11(5):e00166-20.
Other associated proteins
|PF3D7_0302500||cytoadherence linked asexual protein 3.1|
|PF3D7_0905400||high molecular weight rhoptry protein 3|
|PF3D7_1410400||rhoptry-associated protein 1|