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Fuctional and molecular characterization of Trypasomatids spliceosomal proteins


In trypanosomatids, the trans-splicing reaction is responsible for the polycistronic pre-mRNA processing to generate individual genes which contain the 5 end spliced leader (SL) sequence. The ribonucleoproteins (snRNPs) U2, U4/U6, U5 and SL RNP, which consist of snRNAs, 7 Sm proteins (common for all) and specific proteins for each particle, participate in the process. By in vivo production and purification of SmD1 protein-PTP tagged in T. brucei it was possible the identification of 41 proteins that interact with SmD1 particle that which 16 have been annotated as conserved hypothetical. Among them, we already identified a U1-A homolog, a Lsm protein, an U5-specific protein and two proteins that did not present direct binding with the snRNAs analyzed, but probably participate in the processing as splicing factors. This project has as objective the molecular and functional characterization of the other 11 conserved hypothetical proteins to identify new possible chemotherapic targets parasite-specific. For that purpose it will be utilized the PTP-tag technique, primer extension to analyze the binding with snRNAs, RNAi to verify the importance of each protein for the parasite survival and function in the trans-splicing reaction. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
DA SILVA, MARCO TULIO A.; AMBROSIO, DANIELA L.; TREVELIN, CAROLINE C.; WATANABE, TATIANA F.; LAURE, HELEN J.; GREENE, LEWIS J.; ROSA, JOSE C.; VALENTINI, SANDRO R.; CICARELLI, REGINA M. B.. New insights into trypanosomatid U5 small nuclear ribonucleoproteins. Memórias do Instituto Oswaldo Cruz, v. 106, n. 2, p. 130-138, . (07/07476-0, 06/05766-8)
AMBROSIO, DANIELA LUZ; LEE, JU HUCK; PANIGRAHI, ASWINI K.; NGUYEN, TU NGOC; BARRETTO CICARELLI, REGINA MARIA; GUENZL, ARTHUR. Spliceosomal Proteomics in Trypanosoma brucei Reveal New RNA Splicing Factors. Eukaryotic Cell, v. 8, n. 7, p. 990-1000, . (07/07476-0)

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