RESUMO
The 14-3-3 protein is a key player in signal transduction processes in various species. We have previously cloned and expressed the 14-3-3 of Schistosoma mansoni. Using the purified protein we have now raised antibodies against it. A highly specific, affinity-purified antibody preparation was employed for the localization of the 14-3-3 protein in the parasite, by both immunohistochemistry and immunoelectron microscopy. The results demonstrate wide distribution of this protein. It was observed in the female excretory system, the nephridia as well as in the genital systems of both sexes, namely in the vitelline gland of female and in the testis of the male. It is also present in the parenchyma and muscle of both male and female worms. Immunoelectron microscopy demonstrated the presence of immunogold-labelled protein in the tegument, subtegument, muscle, parenchyma and in the female reproductive system, in both the cytoplasm and nucleus of vitelline cells, and oocytes. The possible role of the 14-3-3 protein in the genital organs is discussed.
Assuntos
Schistosoma mansoni/metabolismo , Tirosina 3-Mono-Oxigenase/biossíntese , Proteínas 14-3-3 , Animais , Anticorpos Anti-Helmínticos/biossíntese , Western Blotting , DNA de Helmintos/genética , Feminino , Regulação da Expressão Gênica , Imuno-Histoquímica , Masculino , Microscopia Imunoeletrônica , Reação em Cadeia da Polimerase , Coelhos , Schistosoma mansoni/genética , Schistosoma mansoni/ultraestrutura , Transdução de Sinais/fisiologia , Tirosina 3-Mono-Oxigenase/metabolismoRESUMO
Schistosoma mansoni possesses two isoforms of ferritin, soma and yolk ferritin. The soma ferritin occurs at a low level in most cells of both genders, whereas the yolk ferritin is a female-specific gene product that is expressed at high level in the vitellarium. In higher animals, ferritin mRNA is regulated by iron via the interaction of cytoplasmic binding proteins (IRPs) with a specific sequence element in the 5' untranslated region (UTR) referred to as the iron-responsive element (IRE). Sequence studies of the 5' UTRs, gel retardation assays, and hybridization experiments show that neither ferritin mRNAs of S. mansoni is regulated by an IRE/IRP mechanism. It is suggested that ferritins in schistosomes are controlled only at the transcriptional level.
Assuntos
Ferritinas/genética , Ferro/metabolismo , Processamento Pós-Transcricional do RNA/genética , Schistosoma mansoni/metabolismo , Animais , Sequência de Bases , Northern Blotting , Extratos Celulares/química , Clonagem Molecular , Sondas de DNA/genética , Sondas de DNA/metabolismo , Eletroforese em Gel de Poliacrilamida , Escherichia coli/genética , Ferritinas/classificação , Regulação da Expressão Gênica/genética , Ferro/farmacologia , Dados de Sequência Molecular , Schistosoma mansoni/química , Análise de Sequência , Transcrição Gênica/genéticaRESUMO
Schistosoma mansoni possesses two isoforms of the iron storage protein ferritin, Fer1 and Fer2. At the mRNA level as well as at the protein level, Fer1 is much more abundant than Fer2; females contain an about 15-fold excess of Fer1 compared with males. In contrast, nearly equal amounts of Fer2 occur in both sexes. By electron microscopy we identified ferritin as a component of electron dense membrane-bound bodies in cells of the vitellarium. The mode of formation of these inclusions (as inferred from electron microscopy) and the abundance of phospholipid multilayered membranes suggest that these bodies are of a lysosomal nature. Here we interpret these ferritin-containing inclusions as protein yolk platelets. To date, most of the literature does not contain any hints of the existence of protein yolk in trematodes. The possible function of ferritin in embryonic development is discussed.