title: Molecular characterization of SUMO E2 conjugation enzyme: differential expression profile in Schistosoma mansoni.
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.
authors.affiliation: Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, Morro do Cruzeiro, Ouro Preto, MG, Brazil. robertaverciano@yahoo.com.br
authors.forename: Roberta Verciano
authors.initials: RV
authors.lastname: Pereira
authors.forename: Fernanda J
authors.initials: FJ
authors.lastname: Cabral
authors.forename: Matheus S
authors.initials: MS
authors.lastname: Gomes
authors.forename: Elio H
authors.initials: EH
authors.lastname: Baba
authors.forename: Liana K
authors.initials: LK
authors.lastname: Jannotti-Passos
authors.forename: Omar
authors.initials: O
authors.lastname: Carvalho
authors.forename: Vanderlei
authors.initials: V
authors.lastname: Rodrigues
authors.forename: Robson José Cássia Franco
authors.initials: RJ
authors.lastname: Afonso
authors.forename: William
authors.initials: W
authors.lastname: Castro-Borges
authors.forename: Renata
authors.initials: R
authors.lastname: Guerra-Sá
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D012908:Snails / Q000469:parasitology; D058207:Sumoylation; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000098843:Ubiquitin-Conjugating Enzyme UBC9
chemical_list: D011994:Recombinant Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9