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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2015.tde-27082015-135748
Document
Author
Full name
Tatiane Macedo Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Wunderlich, Gerhard (President)
Azevedo, Mauro Ferreira de
Dutra, Marcos Leoni Gazarini
Title in Portuguese
Estudo do modo de transcrição e expressão dos genes surf de Plasmodium falciparum.
Keywords in Portuguese
Plasmodium falciparum
Antígenos SURFIN
Genes surf
Malária
Transcrição de genes
Abstract in Portuguese
Uma família multigênica, polimórfica, ainda pouco conhecida de P. falciparum é a família SURFIN, codificada por dez genes denominados surf. Para abordar o modo de transcrição desses genes e observar se ocorre switching transcricional, monitoramos a quantidade de transcritos analisando amostras por RT-qPCR ao longo de 40 reinvasões. O gene surf 4 foi claramente detectado em nossos ensaios, apresentando quantidade significativa de transcritos na fase esquizonte. Para analisar a localização e função de SURFIN 4, construímos linhagens transgênicas (por fusão à GFP) e por fusão de um domínio desestabilizador (DD24) que permite de forma controlada, desestabilizar a proteína e analisar a sua importânica. Não houve alteração na viabilidade dos parasitas com SURFIN 4 desestabilizado, no entanto detectamos um aumento significativo de transcritos quando SURFIN 4 foi diminuída. Estes achados apontam para um novo mecanismo de regulação gênica, ao menos para o gene surf 4 onde a proteína SURFIN 4 parece ser capaz de modular a quantidade do seu respectivo transcrito.
Title in English
Studies on the mode of transcription and expression of surf genes from Plasmodium falciparum.
Keywords in English
Plasmodium falciparum
surf genes
Malaria
SURFIN antigen
Transcription profiling
Abstract in English
One smaller family of antigen variant of P. falciparum whose function is still unknown is surf genes. Some members appear in asexual blood stage forms and may be associated also to virulence associated processes. We accessed the transcription of the members of the surf gene family along multiple invasions by real time PCR. Based on the observation of constitutive expression of one surf gene, surf4.1, we created a parasite line which expresses a GFP, conditionally destabilized SURFIN4.1 protein. Upon destabilization of the protein, no differential growth rates were observed. However, when we monitored the transcription of SURFIN4.1 knocked-down parasites compared to their stabilized counterparts, we observed a strong increase in the transcript quantities of surf4.1 and also three other surf genes, pointing to a feedback of the SURFIN protein quantity to either the stability of surf transcripts or the transcriptional activity of surf loci. This type of potential regulation is yet unheard in Plasmodium biology.
 
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Publishing Date
2015-08-28
 
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