<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei"^^ . "Trypanosoma brucei is the causative agent of the African sleeping sickness. Between mammalian hosts it is transmitted by the tsetse fly. Due to the different environments of the hosts the parasite has to adapt its metabolism quickly. T. brucei RNA polymerase II lacks transcriptional control; therefore the control of gene expression is exerted mainly at the level of mRNA stability and translation. RNA stability is influenced by the binding of RNA binding proteins (RBPs), which thereby can play a crucial role in gene expression. This work focused on the characterization of the RNA binding protein RBP10. A polyclonal antibody was raised which showed that RBP10 is only expressed in the BS of the parasite. A knockdown of RBP10 by RNAi in the bloodstream form (BS) of the parasite was lethal after four days. Microarray studies comparing RBP10 knockdown RNA to BS WT RNA revealed a widespread effect on the transcriptome with many BS-specific mRNAs decreased, including many mRNAs encoding proteins involved in glucose metabolism. Further, the effect of the inhibition of glucose uptake by phloretin treatment on the transcriptome was explored and compared to the effect of RBP10 RNAi. The ectopic expression of RBP10-myc in the PC resulted in a defect in proliferation and also in the expression of endogenous RBP10. Microarray studies showed that in the PC the artificial expression of RBP10 lead to a strong increase of many BS specific mRNAs and a simultaneous decrease of PC specific mRNAs. It could also be shown that the forced expression of RBP10 inhibited differentiation of BS to PC trypanosomes. Putative direct RNA targets were identified by IP with subsequent purification of bound RNA and deep-sequencing. However, these results do not overlap with the mRNAs affected after RPB10 RNAi. Also using IP probable protein interaction partners were detected revealing among others RBP29, which is known to be on polysomes, and PABP2. In a sucrose gradient RBP10 was not found in the fractions of the heavy polysomes but could be detected in fractions of the free proteins to the fractions of proteins in trisomes. These findings show that RBP10 is necessary for the expression of many BS specific mRNAs. "^^ . "2011" . . . . . . . . "Martin"^^ . "Wurst"^^ . "Martin Wurst"^^ . . . . . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei (PDF)"^^ . . . "Dissertation_Martin_Wurst.pdf"^^ . . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei (Other)"^^ . . . . . . "indexcodes.txt"^^ . . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei (Other)"^^ . . . . . . "preview.jpg"^^ . . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei (Other)"^^ . . . . . . "medium.jpg"^^ . . . "Characterization of the RNA binding protein RBP10 in Trypanosoma brucei (Other)"^^ . . . . . . "small.jpg"^^ . . "HTML Summary of #12078 \n\nCharacterization of the RNA binding protein RBP10 in Trypanosoma brucei\n\n" . "text/html" . . . "570 Biowissenschaften, Biologie"@de . "570 Life sciences"@en . .