Modificação pós-traducional pendente de SUMO em Schistosoma mansoni: padrão de expressão diferencial durante a transição de cercária a esquistossômulo

AUTOR(ES)
DATA DE PUBLICAÇÃO

2010

RESUMO

Attachment of SUMO to target proteins occur through by similar mechanism to ubiquitination, with sequential action of three enzymes: E1 activating (Aos1-Uba2), E2 conjugation (Ubc9) and E3 ligases (PIAS, Polycomb2 and RanBP2). SUMO is synthesized as inactive precursors that require processing by specific proteases (SENPs). SUMO attachment is a reversible and transient modification. The same enzymes that convert SUMO to its mature form also catalyse the cleavage from their substrates. Reconstitution of SUMOylation pathway in Schistosoma mansoni, based on homology search using avaible databases, shown the existence of nine genes related with SUMOylation: two genes to SUMO (Smsmt3b/c), one to E1 (Smaos1-uba2), one to E2 (Smubc9), two to E3 (Smpias e Smranbp2) and two to SENPs (Smsenp1/7). In this work, the expression levels of these genes was quantified by qRT-PCR and total RNA obtained from cercariae, adult worm and mechanically transformed schistosomula cultivated from 3.5 h to 7 days in vitro (MTS). The results revealed differential gene expression profile to all analyzed genes. We also observed that the transcripts levels were 3 times higher in MTS-3,5h when compared with cercariae and adult worm. The SmUbc9 levels were analysed by Western blot, using specific polyclonal antibodies to S. mansoni protein produced using recombinant protein. The results suggest equivalent levels of SmUbc9 in the same evolutive stages analysed. The SUMOylated substrates profile was detected by Western blot using anti-SUMO. We observed a stage-specific profile of SUMOylated conjugated and an accumulation in initial stages of schistosomula development. These data demonstrate that SUMOylation pathway components are differentially expressed in S. mansoni, suggesting a stage-specific profile during cercariae-schistosomula transition. Whereas this transition is not controlled by real protein synthesis whole results suggest that posttranslational modification SUMO dependent could regulate important biological process during this period of parasite life-cycle.

ASSUNTO(S)

biologia molecular transdução de sinal celular esquistossomose mansônica cercárias

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