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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Estudo funcional da proteína RNF-113A (ZNF183) no spliceossomo em células de mamíferos. / Functional study of RNF113A (ZNF183) in mammalian cells spliceosomes.

Silva, Guilherme Henrique Gatti da 17 March 2017 (has links)
O splicing do pré- RNAm é o processo pelo qual os introns são removidos e os exons são unidos para produzir um RNAm maduro. Esse processo é catalisado por um complexo denominado spliceossomo. RNF113A é uma proteína detectada em complexos pré- e pós-catalíticos. Neste trabalho analisamos a participação de RNF113A no spliceossomo de mamíferos e suas interações com outros componentes do spliceosomo. Nossos resultados mostraram que a super- expressão de RNF113A em células HEK293T aumentam o nível dos snRNAs U1, U2, U5 e U6 e de PRPF19. A imunoprecipitação de RNF113A revelou maior concentração de PRPF19, hBRR2 e SF3b2. Além disso, observamos maior quantidade dos snRNAs U5 e U6 associados à RNF113A. As frações de imunoprecipitação foram analisadas por espectrometria de massas e mostraram interação de RNF113A com proteínas que participam na transcrição, ubiquitinação, maturação do pré-RNAm e splicing. Estes resultados indicam que a associação de RNF113A ao spliceossomo ocorre a partir de suas interações com os snRNAs U5 e U6 e com proteínas relacionadas, como a hBRR2. / Splicing of pre-mRNA is the process by which introns are removed and exons are joined together to produce a mature mRNA competent for translation. It is catalyzed by the complex called spliceosome. RNF113A is a protein detected in pre- and post-catalytic complexes. In this work we analyzed RNF113A participation on spliceosome and its interactions with other spliceosome components. Our results showed that overexpression of RNF113A in HEK293T led to increased levels of U1, U2, U5, U6 snRNAs and PRPF19. RNF113A also associates to PRPF19, hBRR2 and SF3b2, as observed by immunoprecipitation followed by RT-qPCR and western blot. Consistently, U5 and U6 snRNAs were observed in association with RNF113A. Immunoprecipitation fractions were also analyzed by mass spectrometry and results showed interaction of RNF113A with proteins that participate in processes such as transcription, ubiquitination, pre-mRNA maturation and splicing. These results suggest RNF113A associates to spliceosome by interacting with U5 and U6 snRNAs and with related proteins, for example hBRR2.

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