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

Cell Chem Biol. 2022 Sep 15;29(9):1419-1433.e5. doi: 10.1016/j.chembiol.2022.06.001. Epub 2022 Jun 22.

Abstract

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 identified host regulators of cytoskeleton organization, vesicle trafficking, and ER/Golgi stress that regulate LS development. Foci of γ-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 γ-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 recruited to PVM and support parasite development.

Keywords: CENPJ; CRISPR-Cas9; Golgi; MTOC; Plasmodium; genome-wide; liver-stage; malaria; microtubule; vesicular trafficking.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems
  • Humans
  • Liver / metabolism
  • Liver / parasitology
  • Malaria* / metabolism
  • Microtubule-Associated Proteins* / metabolism
  • Microtubules* / metabolism
  • Plasmodium / metabolism
  • Tubulin / metabolism

Substances

  • CENPJ protein, human
  • Microtubule-Associated Proteins
  • Tubulin