Discussion

The leishmaniases, the infections caused by Leishmania parasite species, represent the second largest burden of parasitic disease in the world. Despite this, available treatments are limited; partly due to our incomplete knowledge of the molecular and genetic mechanisms that conduct these parasites through their complex, dixenous lifecycle. Constitutive transcription of polycistrons suggests that post-transcriptional regulators like mRNA-binding proteins (mRBPs) play a prominent role in Leishmania gene regulation.  The Walrad lab has recently isolated the mRNA-bound proteome of the three main lifecycle stages in Leishmania. This dataset has been analysed for candidate mRBPs with a putative involvement in parasite differentiation. By tagging and genetically manipulating levels of prioritised mRBPs, we hope to screen for key regulators which play essential roles in lifecycle progression, infectivity and virulence characterised using a range of bespoke techniques. The fundamental dependence of kinetoplastid parasites on post-transcriptional control also makes Leishmania an ideal model for the study of RNA-binding protein function, providing valuable insight to the broader field of trans-regulators. Immunoprecipitation (IP) and in situ hybridization (FISH) will be used to identify and visualise target mRNAs and protein cofactors that form mRNP ‘regulon’ complexes.  Visualisation of mRNP complexes could provide in vivo data on cellular localisation dynamics in response to key environmental shifts and stress. Isolation of vital gene regulatory pathways could aid in the design of anti-leishmanial therapeutics.