Characterization of nod-like genes in Leishmania (2006)
- USP affiliated authors: ULIANA, SILVIA RENI BORTOLIN - ICB
- USP Schools: ICB
- Subjects: PARASITOLOGIA
- Language: Inglês
- Abstract: Introduction and Objectives: Several proteins of the NOD (nucleotide-binding oligomerization domain) family have been implicated in the intracellular recognition of pathogens and regulation of immune responses. Members of the NOD family in mammals have been associated with the regulation of NF-kB activity and activation of caspases (Annu. Rev. Biochem. 74:355-83, 2005). In the process of studying a genomic region harboring a virulence-related L. amazonensis gene (FEMS Microbiol Lett, 238(1):213-9, 2004), we identified an open reading frame (La17.N3) that presented similarity with the human NOD3 protein. A sequence search in the L. major Friedlin genome allowed the identification of 5 further possible NOD-related genes. One of these genes (LmjF 11.0890) encodes a protein with putative nucleotide-binding domains. The objective of this work is the characterization of 2 of these putative Leishmania NOD-like genes: La17.N3 and LmjF11.0890. Methods and Results: Northern blots indicated that La17.N3 expression is upregulated in L. amazonensis amastigotes, the life cycle stage found in mammalian cells, while LmjF11.0980 seems to be expressed constitutively.The complete nucleotide sequence of La17.N3 was determined and the translated amino acid sequences of La17.N3 and LmjF11.0980 were used to identify orthologous sequences in other Leishmania species. The alignment of these sequences revealed high degrees of similarity. A policlonalantiserum raised against the La17.N3 recombinant protein recognized a 73 kDa protein in total extracts of L. amazonensis and L. major. Constructs for the knock-out and overexpression of the L. amazonensis La17.N3 gene are being concluded. A fragment of 1.2 kb encoding a truncated NOD-like protein LmjF 11.0890 was cloned into a histidine tagged expression vector. The 43-kDa recombinant protein was purified and used to raise specific antibodies. The characterization of this gene and of the encoded protein is under way. Conclusions: The characterization of NOD-like proteins in Leishmania may produce the necessary tools to investigate the participation of these proteins in the macrophage-parasite interaction.
- Título do periódico: Abstracts
- Conference titles: Meeting of the Brazilian Society for Immunology
ABNTFRANCO, F A L; CRISPIM, A F; ULIANA, Silvia Reni Bortolin. Characterization of nod-like genes in Leishmania. Anais.. São Paulo: [s.n.], 2006.
APAFranco, F. A. L., Crispim, A. F., & Uliana, S. R. B. (2006). Characterization of nod-like genes in Leishmania. In Abstracts. São Paulo.
NLMFranco FAL, Crispim AF, Uliana SRB. Characterization of nod-like genes in Leishmania. Abstracts. 2006 ;
VancouverFranco FAL, Crispim AF, Uliana SRB. Characterization of nod-like genes in Leishmania. Abstracts. 2006 ;
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