PRJNA734911
The RNA-binding protein RBP42 regulates cellular energy metabolism in mammalian-infective Trypanosoma brucei
RNA-binding proteins are key players in coordinated post-transcriptional regulation of functionally related genes, defined as RNA regulons. RNA regulons play particularly critical roles in parasitic trypanosomes, which exhibit unregulated co-transcription of long unrelated gene arrays. In this report, we present a systematic analysis of an essential RNA-binding protein, RBP42, in the mammalian-infective bloodstream form of African trypanosome, and show that RBP42 is a key regulator of parasite's central carbon and energy metabolism. Using individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) to identify genome-wide RBP42-RNA interactions, we show that RBP42 preferentially binds within the coding region of mRNAs encoding core metabolic enzymes. Global quantitative transcriptomic and proteomic analyses reveal that loss of RBP42 reduces the abundance of target mRNA-encoded proteins, but not target mRNA, suggesting a positive translational regulatory role of RBP42. To identify direct targets of RBP42, iCLIP was performed using two independent antibodies (anti-RBP42 and BB2), and from extracts prepared from cells following two increasing UV doses, at low (150mJ/cm2) and at high (300mJ/cm2) constant energy. For transcriptomic change mRNA-seq was performed on three biological replicate samples of before (day 0) and three consecutive days after (day 1 -3) RBP42 knockdown.