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Analysis of the interaction between the E74A transcription factor and the regulatory region of the D. melanogaster CGs 7599 and 7336 using trans-activating assays in yeast

Grant number: 12/24005-9
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): February 01, 2013
Effective date (End): January 31, 2015
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Nadia Monesi
Grantee:Danilo Garcia Sanchez
Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil


Our laboratory investigates mechanisms controlling gene expression in metazoans. As a model we employ the DNA puff gene BhC4-1, which is expressed at the end of the larval development of the sciarid Bradysia hygida. Functional assays in the model organism Drosophila melanogaster led to the identification of a salivary gland 129 bp (-186/-58) enhancer in the BhC4-1 promoter. Although the data obtained from gene interaction experiments and bioinformatics analysis indicate that the E74A transcription factor regulates BhC4-1 expression in the salivary gland, it is not possible at the moment to determine whether protein-DNA interactions occur between the transcription factor E74A and the BhC4-1 (-186/-58) salivary gland enhancer. A master's project currently underway in our laboratory aims to determine whether in the context of the yeast one-hybrid system yeast the D. melanogaster E74A transcription factor directly interacts with the BhC4-1 (-186/-58) salivary gland enhancer. This project aims to carry out experiments in the yeast one-hybrid system designed to verify the interaction between the E74A transcription factor and a D. melanogaster 420 bp genomic fragment located between the L71-5 (CG7599) and L71-6 (CG7336) genes, which contains five binding sites for the transcription factor E74A. The binding of E74A to these binding sites has been demonstrated by DNase I protection experiments and functional assays have revealed that these binding sites are required for the proper induction of a L71-6: lacZ transgene by E74A. The results obtained from the development of this project will reveal whether the E74A transcription factor interacts with a DNA fragment, for which the interaction with E74A has previously been demonstrated, in the context of the yeast one-hybrid system, which will be an important positive control for ongoing experiments in the laboratory. (AU)

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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)
FRANK, HENRIQUE OLIVEIRA; SANCHEZ, DANILO GARCIA; OLIVEIRA, LUCAS DE FREITAS; KOBARG, JORG; MONESI, NADIA. The Drosophila melanogaster Eip74EF-PA transcription factor directly binds the sciarid BhC4-1 promoter. GENESIS, v. 55, n. 11, . (12/24005-9)

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