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Molecular characterization of SUMO E2 conjugation enzyme: differential expression profile in Schistosoma mansoni

Author:
Pereira, Roberta Verciano, Cabral, Fernanda J., Gomes, Matheus S., Baba, Elio H., Jannotti-Passos, Liana K., Carvalho, Omar, Rodrigues, Vanderlei, Afonso, Robson José Cássia Franco, Castro-Borges, William, Guerra-Sá, Renata
Source:
Parasitology research 2011 v.109 no.6 pp. 1537-1546
ISSN:
0932-0113
Subject:
Schistosoma mansoni, Western blotting, antibodies, databases, enzymes, gene expression regulation, messenger RNA, post-translational modification, proteins, rats, schistosomula, signal transduction
Abstract:
SUMO-dependent post-translational modification is implicated in a variety of cellular functions including gene expression regulation, nuclear sub-localization, and signal transduction. Conjugation of SUMO to other proteins occurs in a similar process to ubiquitination, which involves three classes of enzymes: an E1 activating, an E2 conjugating, and an E3 target-specific ligase. Ubc9 is the unique SUMO E2 enzyme known to conjugate SUMO to target substrates. Here, we present the molecular characterization of this enzyme and demonstrate its expression profile during the S. mansoni life cycle. We have used bioinformatic approaches to identify the SUMO-conjugating enzyme, the SmUbc9-like protein, in the Schistosoma mansoni databases. Quantitative RT-PCR was employed to measure the transcript levels of SUMO E2 in cercariae, adult worms, and in vitro cultivated schistosomula. Furthermore, recombinant SmUbc9 was expressed using the Gateway system, and antibodies raised in rats were used to measure SmUbc9 protein levels in S. mansoni stages by Western blotting. Our data revealed upregulation of the SmUbc9 transcript in early schistosomula followed by a marked differential gene expression in the other analyzed stages. The protein levels were maintained fairly constant suggesting a post-transcriptional regulation of the SmUbc9 mRNA. Our results show for the first time that S. mansoni employs a functional SUMO E2 enzyme, for the conjugation of the SUMO proteins to its target substrates.
Agid:
427321