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Doctoral Thesis
DOI
10.11606/T.42.2008.tde-03122008-102437
Document
Author
Full name
Fernando Alves de Lima Franco
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Uliana, Silvia Reni Bortolin (President)
Barcinski, Marcello Andre
Tosi, Luiz Ricardo Orsini
Winter, Carlos Eduardo
Yoshida, Nobuko
Title in Portuguese
Caracterização da região genômica META 1 de Leishmania (Leishmania) amazonensis e comparação com a região ortóloga de L. (L.) major.
Keywords in Portuguese
Leishmania
Doenças parasitárias
Genes
Leishmaniose cutânea
Leishmaniose tegumentar difusa
Protozoologia
Abstract in Portuguese
A caracterização de sequências codificadoras presentes nas vizinhanças do gene META 1 permitiu a identificação de alguns genes expressos preferencialmente em estágios infectivos de L. (L.) amazonensis. Um dos genes presentes é regulado de forma distinta, observando-se maior abundância do RNA em formas amastigotas. Este gene foi denominado LaLRR17 por codificar uma proteína contendo em sua região central 6 repetições ricas em leucina (LRR). As LRR são motivos presentes em diversas famílias de proteínas e são responsáveis pela formação de uma estrutura capaz de estabelecer interações protéicas. A região central da proteína LRR17 apresentou similaridade com a porção carboxi-terminal da proteína NOD 3 humana. A proteína LRR17 foi localizada no citosol de macrófagos infectados com L. (L.) amazonensis. Para caracterizar a função da proteína LRR17 foram obtidas linhagens de L. (L.) amazonensis expressoras do gene LmjLRR17. Essas linhagens mutantes foram mais infectivas em macrófagos in vitro quando comparadas com a linhagem selvagem. Avaliamos também o papel das proteínas NOD 1 e NOD 2 na infecção por L. (L.) amazonensis e L. (L.) major para estabelecer a possível relação da proteína LRR17 na interação com essas vias de defesa celular do macrófago.
Title in English
Characterization of the Leishmania (Leishmania) amazonensis genomic region containing the META 1 gene.
Keywords in English
Leishmania
Diffuse cutaneous leishmaniasis
Gene
Leishmaniasis cutaneous
Parasitic diseases
Protozoologia
Abstract in English
The characterization of coding sequences in the vicinity of the META 1 gene allowed the identification of some genes preferentially expressed in L. (L.) amazonensis infective stages. One of the identified transcripts presents a distinct pattern of expression with higher levels of mRNA in amastigotes. This gene was named LaLRR17 since it encodes a 72 kDa protein with 6 leucine-rich repeats (LRR) in its central region. Leucine-rich repeats (LRR) are present in several families of proteins and are responsible for the formation of a structure capable of establishing protein interactions. The central region of the LRR17 protein showed similarity with the carboxyl-terminal portion of the NOD 3 human protein. The LaLRR17 protein is secreted to the cytoplasm of L. (L.) amazonensis-infected macrophages. To characterize the function of the LRR17 protein we obtained strains of L. (L.) amazonensis expressing the LmjLRR17 gene. These mutant strains were more infective to macrophages in vitro when compared with the wild type strain. We also evaluated the role of NOD 1 and NOD 2 proteins in infections with L. (L.) amazonensis and L. (L.) major to investigate a possible role of the LRR17 protein in the interaction with these defense pathways in macrophages.
 
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Publishing Date
2009-04-27
 
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