A genome-wide CRISPR-Cas9 screen identifies CENPJ as a host regulator of altered microtubule organization during Plasmodium liver infection

Prior to initiating symptomatic malaria, a single Plasmodium sporozoite infects a hepatocyte and develops into thousands of merozoites, in part by scavenging host resources, likely delivered by vesicles. Here, we demonstrate that host microtubules (MTs) dynamically reorganize around the developing liver stage (LS) parasite to facilitate vesicular transport to the parasite. Using a genome-wide CRISPR-Cas9 screen, we iden-tified host regulators of cytoskeleton organization, vesicle trafficking, and ER/Golgi stress that regulate LS development. Foci of g-tubulin localized to the parasite periphery; depletion of centromere protein J (CENPJ), a novel regulator identified in the screen, exacerbated this re-localization and increased infection. We demonstrate that the Golgi acts as a non-centrosomal MT organizing center (ncMTOC) by positioning g-tubulin and stimulating MT nucleation at parasite periphery. Together, these data support a model where the Plasmodium LS recruits host Golgi to form MT-mediated conduits along which host organelles are re-cruited to PVM and support parasite development.

Automated summary

This study revealed that host microtubules dynamically reorganize around the developing liver stage parasite to facilitate vesicular transport. Through a genome-wide screen, regulators of cytoskeleton organization, vesicle trafficking, and ER/Golgi stress that regulate liver stage development were identified. The study also found that the Golgi acts as a non-centrosomal microtubule organizing center, promoting microtubule nucleation at the parasite periphery. These findings highlight the importance of host organelles in Plasmodium development.