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Master's Dissertation
DOI
https://doi.org/10.11606/D.17.2018.tde-17042018-143059
Document
Author
Full name
Greicy Kelly Bonifacio Pereira
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2017
Supervisor
Committee
Silva Neto, José Freire da (President)
Koide, Tie
Bertolini, Maria Celia
Silva, Luis Lamberti Pinto da
Title in Portuguese
O papel de uma serina/treonina fosfatase do tipo eucariótico na virulência de Chromobacterium violaceum
Keywords in Portuguese
Chromobacterium violaceum; Fosfatase; Fosforilação; Virulência
Abstract in Portuguese
As proteínas fosfatases desempenham um papel fundamental na modificação pós-traducional reversível de proteínas por fosforilação, ao atuarem na remoção do grupo fosforil estável de resíduos de serina, treonina e tirosina. Embora predominem em eucariotos, as serina/treonina fosfatases (STPs) também existem em bactérias, nas quais atuam controlando diversos aspectos fisiológicos e de virulência. Neste trabalho investigamos o papel das STPs na virulência e fisiologia de Chromobacterium violaceum, uma ?-proteobactéria que atua como um patógeno ocasional de humanos e é amplamente encontrada em água e solos de regiões tropicais e subtropicais. Análises in silico revelaram a presença de quatro STPs no genoma de C. violaceum, sendo que duas possuem apenas o domínio de fosfatase (CV_0881 e CV_3848) e duas possuem um domínio adicional de regulador de resposta (CV_2644 e CV_3505). Mutantes nulos foram construídos para as quatro STPs e todas estas linhagens apresentaram crescimento semelhante ao da linhagem selvagem em meio rico e meio mínimo. As linhagens mutantes ?0881, ?2644 e ?3505 não apresentaram alteração de virulência em camundongos. Os mutantes ?2644 e ?3505 apresentaram menor formação de biofilme, o que sugere papel dessas fosfatases neste processo. Uma caracterização mais aprofundada da linhagem ?3848 por diferentes técnicas de microscopia revelou que este mutante apresenta células de tamanho diminuído, irregularidades no envelope celular e problemas na distribuição do DNA. Estes dados sugerem que CV_3848 tem papel em divisão celular, condensação do material genético e manutenção da parede celular. Nos ensaios de virulência o mutante ?3848 mostrou menor capacidade de causar morte e manter-se no fígado de camundongos, indicando que a STP CV_3848 é importante na patogenicidade de C. violaceum.
Title in English
The role of a eukaryotic-like serine/threonine phosphatase on the virulence of Chromobacterium violaceum
Keywords in English
Chromobacterium violaceum; Phosphatase; Virulence; Phosphorylation
Abstract in English
Protein phosphatases play a key role in reversible post-translational modification of proteins through phosphorylation since they act removing the stable phosphoryl group of serine, threonine and tyrosine residues. Although dominant in eukaryotes, the serine/threonine phosphatases (STPs) also exist in bacteria, in which they act by controlling various physiological and virulence traits. In this work we investigated the role of STPs on the virulence and physiology of Chromobacterium violaceum, a ?-proteobacterium that occasionally acts as a pathogen of humans and it is widely found in water and soil of tropical and subtropical regions. In silico analyzes revealed the presence of four STPs in the genome of C. violaceum, two of which have only the phosphatase domain (CV_0881 and CV_3848) and two that possess an additional domain of response regulator (CV_2644 and CV_3505). Null mutants were constructed for the four STPs and all of them showed similar growth to the wild type strain on rich and minimal medium. The mutant strains ?0881, ?2644 and ?3505 did not show any change in virulence in mice. The ?2644 and ?3505 mutants had lower biofilm formation, suggesting the role of these phosphatases in this process. Further characterization of ?3848 by different microscopy techniques revealed that this mutant presents cells of diminished size, irregularities in the cellular envelope and problem on the DNA distribution. These data suggest that CV_3848 has role in cell division, condensation of the genetic material and cell wall maintenance. In virulence assays the ?3848 mutant showed a lower ability to cause death and to remain in the liver of mice, indicating that the STP CV_3848 is important for the pathogenicity of C. violaceum.
 
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Publishing Date
2018-05-17
 
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