LC3B labeling of the parasitophorous vacuole membrane of Plasmodium berghei liver stage parasites depends on the V‐ATPase and ATG16L1

Abstract The protozoan parasite Plasmodium , the causative agent of malaria, undergoes an obligatory stage of intra‐hepatic development before initiating a blood‐stage infection. Productive invasion of hepatocytes involves the formation of a parasitophorous vacuole (PV) generated by the invagination of the host cell plasma membrane. Surrounded by the PV membrane (PVM), the parasite undergoes extensive replication. During intracellular development in the hepatocyte, the parasites provoke the Plasmodium ‐associated autophagy‐related (PAAR) response. This is characterized by a long‐lasting association of the autophagy marker protein, and ATG8 family member, LC3B with the PVM. LC3B localization at the PVM does not follow the canonical autophagy pathway since upstream events specific to canonical autophagy are dispensable. Here, we describe that LC3B localization at the PVM of Plasmodium parasites requires the V‐ATPase and its interaction with ATG16L1. The WD40 domain of ATG16L1 is crucial for its recruitment to the PVM. Thus, we provide new mechanistic insight into the previously described PAAR response targeting Plasmodium liver stage parasites.