An apicoplast localized GTPase is essential for Toxoplasma gondii survival

The apicoplast is an essential organelle found in Apicomplexa, a large phylum of intracellular eukaryotic pathogens. The apicoplast produces metabolites that are utilized for membrane biogenesis and energy production. A majority of apicoplast resident proteins are encoded by the nuclear genome and are trafficked to the apicoplast, and are referred to as N uclear E ncoded and A picoplast T argeted (NEAT) proteins. In this study, we characterized a NEAT protein named TgBipA, that is a homolog of the highly conserved prokaryotic translational GTPase BipA. BipA is essential for bacterial survival in stress conditions and functions through interactions with the prokaryotic ribosome, although its role is not fully understood. Through genetic knockouts of TgBipA and immunofluorescence imaging we show that loss of TgBipA results in apicoplast genome replication defects, disruption of NEAT trafficking, loss of the apicoplast, and subsequently parasite death. Furthermore, we show through comparative studies that this phenotype closely resembles the delayed death phenomenon observed when inhibiting apicoplast translation. Finally, we show that TgBipA is an active GTPase in vitro, and its GTP hydrolysis activity is critical for its cellular function. Our findings demonstrate TgBipA is a GTPase which has an essential role in apicoplast maintenance, providing new insight into the cellular processes of the organelle.