AP2XI-3 binding sites in Toxoplasma gondii [CUT&Tag]

Toxoplasma proliferation is governed by its flexible but tightly regulated cell division cycle, that remarkably depends on appropriate and temporal transcriptional waves throughout the division cycle although the underlying regulation network is still unclear. Here we reported the transcriptional factor, AP2XI-3, as an essential cell cycle related gene regulator in G1 progression. After AP2XI-3 depletion, tachyzoites growth was obviously arrested in G1 phage and daughter budding progression was also destroyed. Intriguingly, the growth arrest of mutant parasites was also observed in vitro. Combining analysis of RNA sequencing and Cut & TAG revealed the key regulatory role of AP2XI-3 in G1 phage related biomass accumulation and cellular competent. AP2XI-3 controlled and targeted a set of RNA transcription, metabolism or protein biosynthesis related genes, whose dysregulation functionally disrupted cell homeostasis and parasite division. Altogether, our finding demonstrated the functional role of AP2XI-3 in transcriptional controlling of G1 progression and revealed its potential ability as a drug design target. Overall design: Toxoplasma gondii with AP2XI-3 tagged with (mAID-3HA) were collected and processed by CUT&Tag protocol to determine AP2XI-3 binding sites genome-wide in tachyzoites