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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2016.tde-10112016-101050
Document
Author
Full name
Marcelo Machado Paiva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Coltri, Patricia Pereira (President)
Kimura, Edna Teruko
Souza, Gisele Monteiro de
Title in Portuguese
Identificação de proteínas reguladoras do splicing associadas à microRNAs.
Keywords in Portuguese
Splicing
Células cultivadas de tumor
MicroRNA
miR-17-92
RNA
Abstract in Portuguese
Splicing é o processo de remoção de introns e ligação de exons em eucariotos. É realizado pelo spliceossomo, um complexo macromolecular composto por RNAs e mais de cem proteínas. Alguns introns possuem microRNAs, os quais devem ser processados para gerar moléculas maduras. O cluster intrônico miR-17-92 é composto por sete miRNAs que têm sido associados ao desenvolvimento de diferentes tumores em vários tecidos. Neste trabalho o splicing de dois miRNAs deste cluster foi analisado em células HeLa, BCPAP e TPC-I. Os resultados mostraram que introns com miR19a tem o splicing mais eficiente do que aqueles com miR18a em todas as três células analisadas. Além disso, a composição dos spliceossomos foi analisada por espectrometria de massas. Entre as principais proteínas encontradas, destaca-se a presença das hnRNPs, como hnRNP_A1 e hnRNP_A2/B1. Estes resultados são importantes para entender como esses miRNAs são processados, e quais são os principais componentes recrutados em diferentes tipos celulares.
Title in English
Identification of splicing regulatory proteins associated to microRNAs.
Keywords in English
MicroRNA
miR-17-92
RNA
Splicing
Tumour cultivated cells
Abstract in English
Pre-mRNA splicing is the process of intron removal and exon ligation in eukaryotes. It is performed by the spliceosome, a multi-megadalton machinery composed of RNAs and more than a hundred proteins. Intronic miRNAs must be processed from the host gene to generate mature molecules. miR-17-92 is an intronic cluster composed of seven miRNAs which have been associated to the development of different tumors, in several different cells. In this work, we analyzed the splicing of two miRNAs belonging to this cluster in HeLa, BCPAP and TPC-I cells. Interestingly, we observed miR19a is more efficiently spliced than miR18a in all three cells. We also searched for specific proteins that can be involved in their respective splicing process. We observed hnRNP proteins are especially concentrated in spliceosomes assembled in introns containing these miRNAs, based on mass spectrometry data. These results are important to understand how these miRNAs are spliced and matured and also can explain their different expression levels in different cells.
 
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Publishing Date
2016-11-10
 
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