DNA repair genes: functional analysis and evolution

• Grant number: 03/13255-5
• Support Opportunities: Research Projects - Thematic Grants
• Duration: July 01, 2004 - October 31, 2009
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Carlos Frederico Martins Menck
• Grantee: Carlos Frederico Martins Menck
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Most of our research activities are based on the understanding of DNA repair and mutagenesis mechanisms in mammalian cells, as well as other organisms. With the recent enormous amount of data generated by genomic approaches, our project is including now an evolutionary point of view. In this thematic proposal we present 4 different subprojects, and 2 of them are based on studies of DNA repair in mammalian cells, giving priorities to primary cells from human beings. Much of this work will employ the well-succeeded recombinant adenovirus vectors, which were developed in this laboratory. These, carrying DNA repair and DNA repair-related genes will be used to complement cells with human syndromes that present DNA repair deficiencies (such as Xeroderma pigmentosum). The vectors can revert completely cells deficiencies and we are now initiating studies with the perspectives to analyze directly their ability to complement knockout mire for XP genes. Moreover, these vectors will also be used in order to diagnose and identify the genes involved in XP patients as well as in the studies of the dynamics of DNA repair proteins in the cells. Another endpoint to be investigated in mammalian cells is apoptosis induced by DNA damage, as we are trying to decipher the signals that lead the relic to trigger its program of cell death. Adenovirus vectors bearing the photolysis genes are being constructed and they will be tested on their ability to prevent apoptosis in human cells deficient for DNA repair. The involvement of RNA transcription, DNA replication, PARP and cell cycle in signaling for apoptosis in DNA damaged cells are also being investigated. In a different subproject, we are trying to understand the how plant cells defend their genomes from the presence of DNA damage. Two genes that were previously cloned (AtXPB1 e Thi1) have being targeted in this work, and our plan is to identify the roles of the proteins encoded by these genes, in the repair of DNA damage in nuclei (AtXPB) and organelle (Thi1). We also plan to understand how these genes are expressed in plants. Finally, based on the knowledge of the complete genome sequence of Caulobacter crescentus, we intend to identify and investigate the genes involved in the DNA repair of this alpha proteobacteria. Basically, we propose to construct and analyze the phenotype of bacterial mutants on DNA repair related genes. The possibility to synchronize these bacteria may be useful to ascertain the effect of cell cycle on the repair functions in these cells. The differential gene expression along the cells cycle can also be investigated. To our knowledge, this is the first proposal for a genomic approach to investigate DNA repair in bacteria different from the classic Escherichia coli model. Subproject 1) recombinant genetic vectors to study DNA repair in mammalian cells; subproject 2) signaling to apoptosis in mammalian cells with damaged genome; subproject 3) DNA repair genes in plants and functional analysis of the genes AtXPB1 and Thi1; subproject 4) identification and function of genes related to DNA repair in Caulobacter crescentus. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

GODOI, PAULO H. C.; GALHARDO, RODRIGO S.; LUCHE, DOUGLAS D.; VAN SLUYS, MARIE-ANNE; MENCK, CARLOS F. M.; OLIVA, GLAUCIUS. Structure of the thiazole biosynthetic enzyme THI1 from Arabidopsis thaliana. Journal of Biological Chemistry, v. 281, n. 41, p. 30957-30966, Oct. 2006. (03/13255-5)

Filed patent(s) as a result of this research project