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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Insights on a putative aminoacyl-tRNA-protein transferase of Leishmania major

Texto completo
Autor(es):
Sharma, Rohit [1] ; Terrao, Monica Cristina [1] ; Castro, Felipe Freitas [1] ; Breitling, Reinhard [2] ; Faca, Vitor [3] ; Oliveira, Eduardo Brandt [3] ; Cruz, Angela Kaysel [1]
Número total de Autores: 7
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Dept Cell & Mol Biol, Ribeirao Preto Med Sch, Ribeirao Preto, SP - Brazil
[2] Jena Biosci GmbH, Jena - Germany
[3] Univ Sao Paulo, Dept Biochem & Immunol, Ribeirao Preto Med Sch, Ribeirao Preto, SP - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: PLoS One; v. 13, n. 9 SEP 12 2018.
Citações Web of Science: 0
Resumo

The N-end rule pathway leads to regulated proteolysis as an adaptive response to external stress and is ubiquitous from bacteria to mammals. In this study, we investigated a gene coding for a putative core enzyme of this post-translational regulatory pathway in Leishmania major, which may be crucial during cytodifferentiation and the environment adaptive responses of the parasite. Leucyl, phenylalanyl-tRNA protein transferase and arginyl-tRNA protein transferase are key components of this pathway in E. coli and eukaryotes, respectively. They catalyze the specific conjugation of leucine, phenylalanine or arginine to proteins containing exposed N-terminal amino acid residues, which are recognized by the machinery for the targeted proteolysis. Here, we characterized a conserved hypothetical protein coded by the LmjF.21.0725 gene in L. major. In silico analysis suggests that the LmjF.21.0725 protein is highly conserved among species of Leishmania and might belong to the Acyl CoA-N-acyltransferases (NAT) superfamily of proteins. Immunofluorescence cell imaging indicates that the cytosolic localization of the studied protein and the endogenous levels of the protein in promastigotes are barely detectable by western blotting assay. The knockout of the two alleles of LmjF.21.0725 by homologous recombination was only possible in the heterozygous transfectant expressing LmjF.21.0725 as a transgene from a plasmid. Moreover, the kinetics of loss of the plasmid in the absence of drug pressure suggests that maintenance of the gene is essential for promastigote survival. Here, evidence is provided that this putative aminoacyl tRNA-protein transferase is essential for parasite survival. The enzyme activity and corresponding post-translational regulatory pathway are yet to be investigated. (AU)

Processo FAPESP: 13/50219-9 - Investigação sobre fatores e mecanismos do controle de expressão gênica em Leishmania: do papel de modificações pós-traducionais, RNAs não codificadores, cis-elementos e amplificação gênica
Beneficiário:Angela Kaysel Cruz
Modalidade de apoio: Auxílio à Pesquisa - Temático
Processo FAPESP: 11/24086-6 - A "N-end rule" em Leishmania major: caracterização funcional e regulação da expressão gênica de uma Leucil fenilalanil - tRNA-proteina transferase
Beneficiário:Rohit Sharma
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado