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Molecular deconstruction of the cell cycle control in the mouse Y1 adrenocortical cell line

Grant number: 03/02717-8
Support Opportunities:Research Projects - Thematic Grants
Duration: December 01, 2003 - July 31, 2008
Field of knowledge:Biological Sciences - Biochemistry
Principal Investigator:Hugo Aguirre Armelin
Grantee:Hugo Aguirre Armelin
Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated scholarship(s):06/56896-9 - Functions of DUSP 12 and its relation to FGF2 signaling, BE.PQ


For decades (ARMELIN, 1975), our long term goal has been to define the molecular basis of the cell cycle control in mammalian cell lines. In the last 5 years we have been studying the antagonistic interactions between signals initiated in the receptors of FGF2 (Fibroblast Growth Factor 2) and ACTH (Adrenocorticotropic Hormone) or AVP (Arginine Vasopressin) in mouse Y1 adrenocortical cells. FGF2 is a strong mitogen for Y1 cells, whereas ACTH and AVP antagonize FGF2 mitogenic activity. The rationale underlying this approach involved the likely assumption that this experimental study would uncover important reactions comprising the signaling network module that contrais the GO->G1->S transition of the cell cycle in Y1 cells. In fact, the results obtained (LOTFI et al, 2000; LEPIQUE et al, 2000; LOTFI; ARMELIN, 2001; FORTI et al, 2002; SCHWINDT et al, 2003; ROCHA et al, 2003) led us to propose a core for the putative control module we have been searching for. Presently, unpublished results and preliminary observations allow us to amplify the proposed control module to include an extended scheme for the mitogenic pathway triggered by FGF2 to promote the GO->G1->S transition in Y1 cells. This new proposal involves hypotheses whose experimental validation is the aim of this thematic project. The experimental plan of this new thematic project involves multiple methodologies: a) recombinant DNA technology applied to mouse cell lines (cloning sequencing of genes and ESTs, construction of expression vectors, transient and permanent transfection of DNA constructs, etc); b) functional knockout of genes/proteins by interference of RNA (RNAi); c) gene expression analysis by microarrays of cDNA; d) analysis of protein complexes by the yeast two-hybrid method or isolation by affinity followed by resolution in SDS-PAGE and mass spectrometry. (AU)

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