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Doctoral Thesis
DOI
10.11606/T.42.2009.tde-07042009-102906
Document
Author
Full name
Juliana Andreoni Nico
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2009
Supervisor
Committee
Winter, Carlos Eduardo (President)
Ferreira, Henrique Bunselmeyer
Ferro, Emer Suavinho
Ho, Paulo Lee
Yan, Chao Yun Irene
Title in Portuguese
Evidências de redundância funcional entre as pró-hormônio convertases no processamento pós-traducional do precursor da vitelogenina VIT-6 do nematóide Caenorhabditis elegans.
Keywords in Portuguese
Caenorhabditis elegans
Pró-hormônio convertase
Processamento-pós traducional
Redundância funcional
RNAi
Vitelogênese
Abstract in Portuguese
Caenorhabditis elegans possui quatro genes de kpcs (kex2/subtilisin-like proprotein convertases): kpc-1, kpc-2/egl-3, kpc-3/aex-5, kpc-4/bli-4. Em C. elegans, dois dos quatro polipeptídeos de vitelogenina encontrados dentro dos ovócitos, YP115 e YP88, se originam a partir de um precursor polipeptídico (VIT-6) clivado pós-traducionalmente após o motivo RGKR. Nematóides transgênicos foram produzidos com construções repórteres transcricionais de GFP. Foi verificada expressão de kpc-1 tanto em neurônios quanto em células musculares e intestinais. Esses dados, aliados aos dados da literatura para os outros genes kpc de C. elegans, sugerem o envolvimento de KPC-1 no processamento de VIT-6, que é secretada por células intestinais. Ensaios de Western-blot compararam o processamento de VIT-6 em nematóides selvagens, mutantes e knock-down por RNAi para os diferentes genes kpc. A análise de nematóides mutantes e knock-down por RNAi combinado para os outros três genes de convertase de C. elegans confirmou a redundância da atividade dessas enzimas no processamento de VIT-6.
Title in English
Functional redundancy in the post-translational processing of the vitellogenin VIT-6 precursor by Caenorhabditis elegans proprotein convertases.
Keywords in English
Caenorhabditis elegans
Functional redundancy
Post-translational processing
Pro-protein convertase
RNAi
Vitellogenesis
Abstract in English
Four kpc genes are found in the Caenorhabditis elegans genome: (kex2/subtilisin-like proprotein convertases): kpc-1, kpc-2/egl-3, kpc-3/aex-5, kpc-4/bli-4. Two of the four vitellogenin polypeptides, YP115 and YP88, originate from a precursor, VIT-6. VIT-6 is cleaved post-translationally after the RGKR motif. Transgenic worms carrying GFP transcription reporter constructs were produced. Expression of kpc-1 has been localized to neurons as well as muscular and intestinal cells. These data, together with the ones available from the literature for the other kpc genes, suggest the involvement of KPC-1 in the processing of VIT-6, which is secreted from intestinal cells. Western-blot analysis compared the pattern of VIT-6 processing in wild-type, mutants and RNAi-treated worms for the other kpcs. Analysis of worms treated by combined RNAi confirmed the redundancy of KPCs in VIT-6 processing.
 
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Publishing Date
2009-04-08
 
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