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Doctoral Thesis
DOI
10.11606/T.42.2008.tde-22092008-130133
Document
Author
Full name
Regina Célia Pereira Marques
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Menck, Carlos Frederico Martins (President)
Baldini, Regina Lúcia
Ferreira, Rita de Cassia Cafe
Italiani, Valeria Cristina da Silva
Lima, Lucymara Fassarela Agnez
Title in Portuguese
Identificação de genes de reparo de DNA em Caulobacter crescentus através da seleção de clones sensíveis a agentes genotóxicos.
Keywords in Portuguese
Caulobacter crescentus
Agentes genotóxicos
Genômica funcional
Lesão no DNA
Reparação de DNA
Resposta a estresses
Abstract in Portuguese
Em estudos de proteção ao genoma contendo lesões de C. crescentus, foi feita uma varredura em uma biblioteca clones mutados pelo transposon Tn5 e sensíveis à luz UV-B e MMS. Dos 249 clones selecionados conseguimos identificar mutações em 102 genes, distribuídos em dez categorias funcionais, incluindo metabolismo de DNA. Além de genes já descritos como atuantes na defesa de genoma a lesões, os dados ainda adicionam funções potenciais para outros genes. Investigação mais detalhada indica que vários dos genes identificados podem afetar o estresse e ciclo celular, podendo estar relacionados a respostas do tipo pontos de checagem. Além disso, verificamos que mutantes para genes envolvidos em reparo excisão de nucleotídeos são mais sensíveis à H2O2, indicando que nessa bactéria, este sistema de reparo atua também em lesões oxidativas no DNA. Os dados obtidos são pioneiros no estabelecimento de funções para vários genes de C. crescentus e de outras alfa-proteobactérias.
Title in English
Identification DNA repair related genes in Caulobacter crescentus through the identification of mutations affecting sensitivity to genotoxic agents.
Keywords in English
Caulobacter crescentus
DNA damage
DNA repair
Functional genomic
Genotoxics agents
Stress response
Abstract in English
This work aimed to identify genes related to DNA damage protection in C. crescentus, by means of screening a Tn5 -mutated library for clones sensitive to UV-B light and MMS. Out of 249 selected clones, we were able to identify mutations in 102 genes, classified in ten different functional categories, including DNA metabolism. In addition to genes already described as playing a role in genome defense to lesions, the results provide new potential functions to several other genes. More detailed investigation indicates also that the mutations in some of these genes may also affect cell stress and cell cycle, being potentially involved in check-point responses. Moreover, mutants for nucleotide excision repair genes were also found to be sensitive to H2O2, indicating that this DNA repair system also acts in DNA oxidative damage. These data are the first establishing a role in genome protection for several genes in C. crescentus, as well as other alpha-proteobacteria.
 
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Publishing Date
2008-09-24
 
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