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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2011.tde-20012012-150835
Document
Author
Full name
Alessandro Aparecido Rodrigues da Silva
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Uliana, Silvia Reni Bortolin (President)
Carboni, Beatriz Simonsen Stolf
Mestriner, Clara Lúcia Barbieri
Tosi, Luiz Ricardo Orsini
Wunderlich, Gerhard
Title in Portuguese
Análise funcional da proteína LRR17, rica em repetições de leucina e secretada por Leishmania (Viannia) braziliensis e L. (Leishmania) amazonensis.
Keywords in Portuguese
Leishmania brasiliensis
Leishmania
Genomas (análise)
Proteínas (análise)
Secreção (análise)
Abstract in Portuguese
As repetições ricas em leucina (LRR) são domínios presentes em diversas famílias de proteínas com diferentes funções, sendo responsáveis pela formação de uma estrutura capaz de estabelecer interações protéicas. Em decorrência do projeto de caracterização de um segmento do cromossomo 17 de L. amazonensis, identificamos um gene que codifica uma proteína de 72 kDa, contendo em sua região central, seis motivos LRR. Genes ortólogos estão presentes nos genomas de L. major e L. braziliensis. Observamos que o gene LRR17 é regulado de forma distinta ao longo dos ciclos biológicos de L. braziliensis e L. amazonensis. A proteína LRR17 de L. braziliensis e secretada tanto nos estágios promastigota como amastigota. Identificamos também a secreção da proteína LRR17 em promastigotas de L. amazonensis. Obtivemos mutantes hiperexpressores da proteína LRR17 em L. braziliensis e L. amazonensis. As linhagens mutantes foram mais infectivas em infecções de macrófagos in vitro quando comparadas com a linhagem selvagem. A proteína LRR17 parece estar envolvida no processo de invasão do parasita em infecções in vitro e no estabelecimento da infecção da forma amastigota de Leishmania.
Title in English
Functional analysis of the LRR17 protein, rich in leucine repeats and secreted by Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis.
Keywords in English
Leishmania brasiliensis
Leishmania
Genome (analysis)
Protein (analysis)
Secretion (analysis)
Abstract in English
Leucine-rich repeats (LRRs) are versatile binding motifs found in a variety of proteins involved in protein-protein interactions. The LaLRR17 gene, identified initially in the L. amazonensis genome, encodes a protein with 6 LRR in its central region, that is secreted to the cytoplasm of L. amazonensis-infected macrophages. An orthologue to LaLRR17 was identified in L. braziliensis chromosome 17. LRR17 gene expression is regulated differentially during the life cycle of L. braziliensis and L. amazonensis. The LbLRR17 protein is secreted in L. braziliensis promastigotes and amastigotes. To characterize the function of the LRR17 protein we obtained transgenic parasite lines of L. amazonensis overexpressing the LaLRR17 gene and of L. braziliensis overexpressing the LbLRR17 gene. The mutants were more infective to macrophages in vitro when compared with the wild type strains, indicating that the LRR17 protein may interact with macrophage molecules, modulating the cellular response to increase parasite survival.
 
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Publishing Date
2012-03-20
 
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