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Role of phosphorylatable residues in the transcriptional activity of the neural protein cScrt2

Grant number: 14/04451-0
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): May 01, 2014
Effective date (End): October 31, 2014
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Chao Yun Irene Yan
Grantee:Mariana Soares Fogo
Home Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil


The family SCRATCH of transcription factors belongs to the superfamily Snail, whose members present a carboxyl-terminal region conserved composed by from 4 to 6 zinc-fingers domains, responsible for the DNA binding on the consensus site E-box. The amino-terminal region, less conserved, contains the SNAG domain and seems to be responsible for the recruitment of transcriptional repression machinery. Studies in different animal models show that Scratch has an important role in neural development. In Gallus gallus, the Scratch homologous (cScratch2) is expressed only in the neural tube and overlaps factors expressed in the beginning of the neural differentiation which also recognizes E-box sites, indicating the occurrence of competition and, possibly, modulation between these factors. These data together suggest that cScrt2 acts on the early neural development controlling or modulating the transcription of genes that contain E-box elements in their cis-regulatory regions. In silico analysis show three possible phosphorylation targets (Tyrosine77, Serine78 and Serine82) that can have important role in cScrt2 activity. This way, the aim of this project is to investigate the importance of two residues (Tyrosine77 and Serine78) in the transcriptional activity of cScrt2. For this, HEK293T cells will be co-transfected with plasmids containing cScrt2 (wild type or mutated) and luciferase, which will make possible to compare indirectly the transcriptional activity of cScrt2 (wild type or mutated) through luciferase activity measurement.