Protein-Protein Interactions Associated with Trypanosoma Brucei Splicing Factor SF3A60
Dweni, Celestine Khayell
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African Trypanosomiasis is caused by Trypanosoma brucei, a flagellated unicellular lower eukaryote of the family Kinetoplastida. Currently, there is no vaccine against the disease and the available anti-parasitic regimen is costly and ineffective hence the quest for alternative molecular targets for chemotherapeutic intervention. Trypanosoma brucei processes its protein coding mRNAs via splice leader (SL) trans-splicing which is a key difference between the parasites and the mammalian hosts and is carried out by the spliceosome. In order to exploit the existence of such specific molecular mechanisms in the quest for novel drug targets, it is crucial to identify and characterize protein factors and their interactions within a targeted trypanosome molecular complex. The SF3a60 is an essential component of the spliceosome which recruits the SF3a120 into the U2 particle during the formation of the spliceosome. In a pilot study, genome wide Yeast two-Hybrid screening using the SF3a60 as bait, identified 56 interacting partners making it a potential protein hub within the trypanosome transspliceosome. This study was aimed at identifying and characterizing SF3a60 associated proteins and their interactions. Potential trans-spliceosomal factors were selected based on their sequence specific signatures. The NP-40 lysis procedure and indirect immunofluorescence assays were used to determine the sub-cellular localization of the trans-spliceosomal proteins. Homology prediction using trypanosome homologues of the SF3a60 protein preys in .the Saccharomyces Genome database was used to identify protein-protein interactions. Out of an initial pool of 56 SF3a60 protein preys, eleven could be involved in the trypanosome splicing process. In addition, six of them had tetracopeptide domains and their sub-cellular localization revealed both nuclear and cytoplasmic distribution. Two protein-protein interactions were predicted between Tb927.3.3220 and Tb927.7.4080 and also Tbl1.01.7710 and Tb927.7.1560. These interactions' were added to the existing A complex to form a preliminary A complex interactome. These proteins and their interactions pose as potential targets for a rational drug design.