Oral Presentation Inaugural Australian Ubiquitin Summit 2025

A CRISPR screen reveals a GID/CTLH-like E3 ubiquitin ligase that controls Toxoplasma development into its latent form (129509)

Alex Uboldi 1 , Sachin Khurana 1 , Sai Lekkala-Lethakula 1 , Amber Simonpietri 1 , Karan Singh 1 , Vinzenz Hofferek 2 , Ushma Ruparel 1 , Lachlan Whitehead 1 , Kelly Rogers 1 , Alexandra Garnham 1 , Nichollas Scott 3 , Nicholas Katris 2 , Malcolm McConville 2 , David Komander 1 , Simon Cobbold 1 , Chris Tonkin 1
  1. WEHI, Melbourne, VIC, Australia
  2. Department of Biochemistry and Pharmacology, The University of Melbourne, Melbourne, Victoria, Australia
  3. Department of Microbiology and Immunology, The University of Melbourne, Melbourne, Victoria, Australia

Toxoplasma and other Apicomplexan parasites, switch between different developmental stages to persist in and transmit between hosts. Toxoplasma can alternate between systemic tachyzoites and encysted bradyzoite forms found in the CNS and muscle tissues. How parasites sense these tissue types and trigger differentiation remains largely unknown. We show that Toxoplasma differentiation into latent forms is induced under metabolic conditions that mimic CNS and muscle tissue and using a series of in vitro and in vivo CRISPR screen identify parasite genes required for this process. From ~25 genes we find as being important for differentiation we find almost every component of a E3 ubiquitin ligase complex orthologous to the glucose induced degradation deficient (GID) complex in yeast and CTLH complex in humans. We show that the makeup of this complex and show that it is localised in the cytoplasm and nucleus of Toxoplasma. We show that TgGID likely regulates translational repression of a key transcription factor required for differentiation, BFD1, through its 3’ utr. Overall, this work provides important new insight into how these divergent parasites sense different host cell niches and trigger stage conversion through a ubiquitination-dependent program.