| 2023 |
BFD2 is a cytosolic RNA-binding protein of the CCCH-type zinc-finger family that is transcriptionally activated by BFD1. BFD2 interacts with the BFD1 transcript under stress conditions, and deletion of BFD2 reduces BFD1 protein levels without affecting BFD1 mRNA abundance, indicating BFD2 acts as a post-transcriptional regulator of BFD1 translation. This establishes a positive feedback loop in which BFD1 drives BFD2 transcription and BFD2 promotes BFD1 translation, enforcing the chronic-stage gene-expression programme. |
Genetic knockout (ΔBFD2), RNA binding assay (BFD2 co-immunoprecipitation with BFD1 mRNA under stress), western blot and mRNA quantification to separate protein vs mRNA levels, in vitro and in vivo differentiation assays |
Nature microbiology |
High |
37081202
|
| 2023 |
Parasites lacking BFD2 fail to induce BFD1 protein and are consequently unable to fully differentiate into bradyzoites either in culture or in mice, demonstrating BFD2 is required for stage conversion. |
Genetic knockout (ΔBFD2), in vitro differentiation assays, in vivo mouse infection with cyst burden assessment |
Nature microbiology |
High |
37081202
|
| 2024 |
BFD2/ROCY1 is necessary for cap-independent translation of BFD1 through direct binding to the 5'-leader of BFD1 mRNA. This was demonstrated by showing that translational control of BFD1 persists when ribosome assembly near the 5'-cap is blocked (by a 5'-proximal stem-loop insertion or knockdown of cap-binding protein eIF4E1), and that BFD2 knockdown abolishes this cap-independent translation. |
Biochemical and molecular approaches: 5'-leader reporter assays, 5'-proximal stem-loop insertion, eIF4E1 knockdown, BFD2 knockdown, cap-independent translation assays |
The Journal of biological chemistry |
High |
39542243
|
| 2024 |
Translation initiation factor eIF1.2 is required upstream of BFD2 for stress-induced differentiation; Δeif1.2 parasites fail to upregulate BFD1 and BFD2 during stress, and forced expression of BFD2 (or BFD1) significantly restores differentiation in Δeif1.2 parasites, placing BFD2 downstream of eIF1.2 in the differentiation pathway. |
Genetic ablation (Δeif1.2), RNA sequencing, ribosome profiling, forced overexpression of BFD2, in vitro and in vivo cyst formation assays |
Nature communications |
High |
37961607 38782906
|
| 2023 |
ROCY1 (the same protein as BFD2) encodes an RNA-binding protein whose RNA-binding domains are required to mediate bradyzoite development; it has a preference for 3' regulatory regions of hundreds of T. gondii transcripts, and is sufficient for bradyzoite formation in vitro and in vivo. |
Genetic knockout (ΔROCY1), domain mutation of RNA-binding domains, RIP-seq or binding preference assays, in vitro and in vivo bradyzoite/cyst formation assays |
Nature communications |
High |
37770433
|
| 2025 |
BFD2 knockout (ME49Δbfd2) reduces parasite proliferation and plaque formation in vitro, and in vivo Δbfd2 parasites induce reduced inflammatory cytokine expression (IL-1β, IFN-γ, TNF-α) in infected BV2 cells and reduce brain inflammation and neuronal apoptosis specifically during acute infection, linking BFD2 to regulation of acute-stage host-parasite interactions. |
CRISPR/Cas9-mediated BFD2 knockout, RNA sequencing of infected BV2 cells, western blot, qPCR, ELISA, in vivo mouse infection with behavioural testing (elevated plus maze, open field test) |
PLoS neglected tropical diseases |
Medium |
40906736
|