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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

Caracterização da interação entre o metilossomo e o nucleocapsídeo do vírus respiratório sincicial humano. / Characterization of humam respiratory syncytial virus nucleoprotein and methylosome interaction.

Ogawa, Juliana Kaori 07 December 2016 (has links)
Neste projeto caracterizamos a interação da nucleoproteína viral (N) do Vírus Respiratório Sincicial Humano (HRSV) com as proteínas PRMT5 e WDR77, que constituem o metilosomo celular. Confirmamos que essa interação ocorre através de co-imunoprecipitação em células humanas, e de interação in vitro dessas proteínas purificadas. Demonstramos a co-localização dessas proteínas na célula através de microscopias de imunofluorescência e confocal. Inibindo ou aumentando a expressão de PRMT5 não observamos impacto na replicação viral. Verificamos que ocorre metilação em N tanto em resíduos de argininas como de lisinas, com anticorpos específicos para essas modificações, e por espectrometria de massas, indicando significado funcional. Com essa evidência testamos o efeito de inibidores de metilação e demetilação, em argininas e lisinas. Obtivemos efeito inibitório significativo da replicação do HRSV com um inibidor de metilação de lisina, UNC0646, indicando que a interação N-metilossomo tem potencial como alvo terapêutico contra HRSV. / In this project we had as objective to characterize the interaction observed previously in the laboratory of viral nucleoprotein (N) with PRMT5 and WDR77 proteins that constitute the cell metilosome. We confirmed that this interaction occurs through co-imunoprecipitation in human cells and in vitro interaction of these purified proteins. We also demonstrated the co-localization of these proteins in inclusion bodies, by immunofluorescence and confocal microscopy. Inhibiting or enhancing PRMT5 expression we didnt see effect on viral replication. Our results show that methylation occurs in both arginine and lysine residues, through reactivity with antibodies specific to these modifications, and analysis by mass spectrometry. We tested the effect of arginine and lysine methylation and de-methylation inhibitors in viral replication. We obtained significant inhibitory effect on HRSV replication with a lysine methylation inhibitor, UNC0646, indicating that N-metilossome interaction has the potential to be exploited as a therapeutic target in developing antiviral drugs.

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