Chemical Rescue of Malaria Parasites Lacking an Apicoplast Defines Organelle Function in Blood-Stage Plasmodium falciparum.docVIP

ChemicalRescueofMalariaParasitesLackingan ApicoplastDefinesOrganelleFunctioninBlood-Stage Plasmodiumfalciparum EllenYeh1*,JosephL.DeRisi2,3* 1DepartmentofPathology,StanfordMedicalSchool,Stanford,California,UnitedStatesofAmerica,2DepartmentofBiochemistryandBiophysics,UniversityofCalifornia, SanFrancisco,California,UnitedStatesofAmerica,3HowardHughesMedicalInstitute,ChevyChase,Maryland,UnitedStatesofAmerica Abstract Plasmodium spp parasites harbor an unusual plastid organelle called the apicoplast. Due to its prokaryotic origin and essentialfunction,theapicoplastisakeytargetfordevelopmentofnewanti-malarials.Over500proteinsarepredictedto localizetothisorganelleandseveralprokaryoticbiochemicalpathwayshavebeenannotated,yettheessentialroleofthe apicoplast during human infection remains a mystery. Previous work showed that treatment with fosmidomycin, an inhibitor of non-mevalonate isoprenoid precursor biosynthesis in the apicoplast, inhibits the growth of blood-stage P. falciparum. Herein, we demonstrate that fosmidomycin inhibition can be chemically rescued by supplementation with isopentenylpyrophosphate(IPP),thepathwayproduct.Surprisingly,IPPsupplementationalsocompletelyreversesdeath following treatment with antibiotics that cause loss of the apicoplast. We show that antibiotic-treated parasites rescued with IPP over multiple cycles specifically lose their apicoplast genome and fail to process or localize organelle proteins, rendering them functionally apicoplast-minus. Despite the loss of this essential organelle, these apicoplast-minus auxotrophs can be grown indefinitely in asexual blood stage culture but are entirely dependent on exogenous IPP for survival. These findings indicate that isoprenoid precursor biosynthesis is the only essential function of the apicoplast duringblood-stagegrowth.Moreover,apicoplast-minusP.falciparumstrainswillbeapowerfultoolforfurtherinvestigation ofapicoplastbiologyaswellasdrugandvaccinedevelopment. Citation: Yeh E, DeRisi JL (2011)