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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2015.tde-04092015-142744
Document
Author
Full name
Aline Sampaio Pinto
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Fernandes, Andrea Balan (President)
Aniz, Patricia Antonia Estima Abreu de
Barros, Mário Henrique de
Dias, Marcio Vinícius Bertacine
Moreno, Ana Carolina Ramos
Title in Portuguese
Caracterização estrutural e análises funcionais das proteínas periplasmáticas NrtT e PotF de transportadores do tipo ABC de Xanthomonas axonopodis pv. citri.
Keywords in Portuguese
Alcano sulfonatos
Fitopatógenos
Proteínas de transporte
SAXS
Abstract in Portuguese
Xanthomonas citri (X. citri), causador do cancro cítrico, afeta muitas áreas de cultivo de citros com impacto comercial. Transportadores ABC foram relatados como essenciais para a patogênese de X. citri. O gene nrtT codifica a proteína periplasmática responsável pelo suposto transporte de nitrato/nitrito/taurina. Neste trabalho, mostramos que a deleção de nrtT não afeta o crescimento de X. citri, porém reduz e atrasa os sintomas do cancro durante a infecção em folhas de citros. Observamos redução na produção de goma xantana e capacidade de aderência na linhagem mutante. A proteína NrtT foi expressa monomérica e monodispersa. Foram observadas alterações na estrutura secundária e aumento da estabilidade térmica de NrtT na presença de MOPS, indicando ser este um possível ligante de NrtT. A proteína PotF descrita como ligadora de putrescina não sofre alteração significativa na estabilidade térmica na presença de putrescina, entretanto, dados de SAXS mostram alterações na estrutura possivelmente decorrentes da ligação com putrescina.
Title in English
Structural characterization and functional analysis of the NrtT and PotF periplasmic proteins of Xanthomonas axonopodis pv. citri ABC transporter.
Keywords in English
Alkanesulfonates
Phytopathogens
SAXS
Transporter proteins
Abstract in English
Xanthomonas citri (X. citri), which causes citrus canker, affects many citrus growing areas with commercial impact. ABC transporters have been reported as essential for the pathogenesis of X. citri. The nrtT gene encodes the periplasmic protein responsible for the alleged transport of nitrate/nitrite/taurine. In this work, we show that nrtT deletion does not affect the growth of X. citri, but delays and reduces the symptoms of cancer during infection of citrus leaves. We observed a reduction in the production of xanthan gum and adhesion capacity in the mutant strain. The NrtT protein was expressed monomeric and monodisperse. There were changes in the secondary structure and increased thermal stability NrtT in the presence of MOPS, indicating that this was a possible binder NrtT. The PotF protein described as putrescine-binding is not significantly altered thermal stability in the presence of putrescine, however, SAXS data shows changes in the structure possibly resulting from connection with putrescine.
 
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Publishing Date
2015-09-04
 
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