RESUMO
The myosin regulatory light chain (RLC) proteins play an important role in cellular processes, especially in muscle contraction. The planarian intestine is a fascinating system for studying the organogenesis during regeneration. In this paper, A homolog gene of Rlc, DjRlc, was identified and characterized in Dugesia japonica. The DjRlc sequence analysis revealed that it contains an opening reading frame encoding a putative protein of 175 amino acids with functionally domains that are highly conserved, including an EF-Hand motif and Ca2+ binding sites. Whole mount in situ hybridization showed that DjRlc is predominantly expressed in the intestine of intact and regenerating planarians. The cross sections of planarians revealed that the DjRlc distributes in the muscle of intact planarians. Knockdown of RNA interference of DjRlc by dsRNA-DjRlc affected the intestinal morphology, causing distinct defects in branching morphogenesis. These finding suggest that DjRlc is required for intestinal regeneration.
Assuntos
Intestinos/fisiologia , Cadeias Leves de Miosina/genética , Planárias/genética , Regeneração/genética , Sequência de Aminoácidos , Aminoácidos/genética , Animais , Sequência de Bases , Sítios de Ligação/genética , Cálcio/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Proteínas de Helminto/genética , Morfogênese/genética , Fases de Leitura Aberta/genética , Filogenia , Interferência de RNA/fisiologia , Alinhamento de SequênciaRESUMO
14-3-3 proteins are a family of highly conserved acidic proteins that regulate cellular processes. They act as a kind of important signaling molecules taking part in many crucial decisions throughout the development process. We have isolated and characterized two members of the 14-3-3 family, namely, Dj14-3-3 α and Dj14-3-3 ζ in the planarian Dugesia japonica. The Dj14-3-3 α and ζ genes encode polypeptides of 260 and 255 amino acids respectively. We have proved that the Dj14-3-3 α and ζ genes were especially expressed in the pharynx in adult and regenerating planarians by in situ hybridization and they were not involved in regeneration process. Besides, Dj14-3-3 α and ζ genes can compensate each other in planarians by RNA interference. The Dj14-3-3 α and ζ were significantly up-regulated expression when planarians were stimulated with the pathogen-associated molecular patterns including lipopolysaccharide (LPS), peptidoglycan (PGN), ß-Glu and Poly (I:C), indicating that the Dj14-3-3 α and ζ may be involved in the immune responses.