We applied the FKBP destabilization domain (DD) technique that allows modulating expression levels through the stabilizing compound Shield-1 and generated a clonal parasite line expressing endogenous PfHP1 as a C-terminally tagged GFP-DD fusion (3D7/HP1ON) (B) Expression and localization of PfHP1 in 3D7/HP1ON, 3D7/HP1OFF, and 3D7/HP1ctrl parasites by live fluorescence microscopy (images taken 12 hr after removal of Shield-1). Live-cell imaging revealed the expected perinuclear localization of tagged PfHP1 in 3D7/HP1ON and 3D7/HP1ctrl parasites throughout the IDC, whereas in 3D7/HP1OFF parasites PfHP1 was undetectable 12 hr after Shield-1 withdrawal. (C) Expression and localization of PfHP1 in 3D7/HP1ON and 3D7/HP1OFF parasites by IFA and western blot (Shield-1 removal at 4–12 hpi). After Shield-1 removal at 4–12 hpi, PfHP1 was still detectable but reduced in late ring stages (16–24 hpi) and early schizonts (32–40 hpi) and localized diffusely to the nucleoplasm and cytoplasm.
Brancucci NM, Bertschi NL, Zhu L, Niederwieser I, Chin WH, Wampfler R, Freymond C, Rottmann M, Felger I, Bozdech Z, Voss TS. Heterochromatin protein 1 secures survival and transmission of malaria parasites. Cell Host Microbe. 2014 16(2):165-76.