Article
Live-cell imaging of mitogen-activated protein kinase localization in Plasmodium berghei liver stage parasites
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Published: | January 29, 2014 |
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Mitogen-activated protein kinases (MAPKs) mediate a variety of cellular functions like stress response, proliferation and differentiation in eukaryotic cells. Two MAPK homologs designated as MAPK1 and MAPK2 have been identified in Plasmodium.
For Plasmodium berghei liver stage parasites, transcription of both genes was confirmed by RT-PCR. GFP-tagged PbMAPK1, expressed either under control of the strong constitutive EEF1a promoter or the endogenous pbmapk1 promoter, displayed a stage-dependant and dynamic subcellular localization pattern during liver stage development. In the early liver schizont, PbMAPK1 localized inside the parasite’s nuclei, whereas a distinct localization of PbMAPK1 in comma/ring-shaped structures in proximity to the parasite’s nuclei and the invaginating parasite membrane was observed during late liver stage development. In contrast, GFP-tagged PbMAPK2 showed a constitutive association with the dividing parasite nuclei. PbMAPK1 knock-out parasites displayed normal host cell invasion and completion of in vitro liver stage development.
To further characterize the PbMAPK1 localization in late liver stage parasites, subcellular compartments putatively related to the PbMAPK1 localization were visualized by live-cell imaging of transgenic parasites expressing GFP-tagged fusion proteins targeting phosphoinositides and structures involved in microtubular organization.