Protein-Protein Interactions Associated with Trypanosoma Brucei Splicing Factor SF3A60
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Date
2013-10-16
Authors
Dweni, Celestine Khayell
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Abstract
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.
Description
Department of Biochemistry and Biotechnology, 86p. The QL 368 .K8D86 2013
Keywords
Trypanosoma brucei --Kenya