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Doctoral Thesis
DOI
https://doi.org/10.11606/T.46.2014.tde-22012015-145300
Document
Author
Full name
Marcela Bach Prieto
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2014
Supervisor
Committee
Oliveira, Carla Columbano de (President)
Barros, Mário Henrique de
Farah, Shaker Chuck
Haddad, Luciana Amaral
Silva, Aline Maria da
Title in Portuguese
Caracterização funcional das proteínas Nop17p e Rsa1p de Saccharomyces cerevisiae
Keywords in Portuguese
Expressão gênica
Nop17p
R2TP
Rsa1p
SnoRNA U3
SnoRNPs de box C/D
Abstract in Portuguese
Nop17p e Rsa1p são proteínas nucleolares em Saccharomyces cerevisiae, as quais foram identificadas pela sua associação a dois complexos celulares: os snoRNPs de box C/D, através de interação com as subunidades Nop58p e Snu13p, respectivamente, e o R2TP/Hsp90p. Nop17p parece ser responsável por direcionar a chaperona Hsp90p durante a montagem dos snoRNPs, e a associação de Rsa1p a estes complexos ainda não tem uma função estabelecida. Neste trabalho, nós mostramos que a ausência de ambas as proteínas afetam a estabilidade da proteína Nop58p dos snoRNPs e afetam a localização do snoRNA U3. Em relação à ordem de interação das proteínas do core de snoRNps de box C/D, Nop17p associa-se de maneira transiente a Nop1p/Snu13p, seguida da ligação de Nop58p ao complexo. Quanto à rede de interação do R2TP, obtivemos o mutante Nop17(N307S), que não mais interage com Tah1p. Este mutante interage com a subunidade Rvb1p do R2TP, mas não se associa com outras proteínas parceiras de Nop17p(WT). Apesar da importância da interação Nop17p-Tah1p, sua interrupção não afeta o crescimento celular, o que sugere a possibilidade de outro fator estar envolvido na associação entre Nop17p e Hsp90p.
Title in English
Functional characterization of the Saccharomyces cerevisiae proteins Nop17p and Rsa1p
Keywords in English
Box C/D snoRNPs
Gene expression
Nop17p
R2TP
Rsa1p
U3 snoRNA
Abstract in English
Nop17p and Rsa1p are Saccharomyces cerevisiae nucleolar proteins, which were identified for its association with two cellular complexes: box C/D snoRNPs, through interaction with the core subunits Nop58p and Snu13p respectively, and the R2TP/Hsp90p. Nop17p seems to be responsible for directing Hsp90p to the assembly of snoRNPs. The Rsa1p association to these complexes still have no defined function. In this work, we showed that both proteins absence affect Nop58p stability and causes a mislocalization of the U3 snoRNA. Relativel to the order of assembly of the box C/D snoRNPs core proteins, Nop17p associates transiently with Nop1p/Snu13p, followed by the Nop58p joining to the complex. To study in more detail the protein interactions within the R2TP complex, we obtained the Nop17(N307S) mutant, which no longer interacts withTah1p, but still interacts withRvb1p, another R2TP subunit. Nop17(N307S) does not interact with other Nop17p(WT) partners. Despite the importance of the Nop17p-Tah1p association, the disruption of this interaction does not affect cell growth, suggesting the involvement of a second factor on the Nop17p and Hsp90p association.
 
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Publishing Date
2015-03-18
 
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