IDENTIFICATION OF YEAST PKA SUBUNITS TRANSCRIPTION REGULATORS USING TWO-COLOR CELL ARRAY SCREENING

PAUTASSO, C; CAÑONERO, L; ZAREMBERG, V; ROSSI, S

Mendoza

Congreso; XLVIII Reunión Anual de la Sociedad Argentina de Bioquímica y Biología Molecular (SAIB); 2012

SAIB

Transcriptional regulation is a key mechanism that controls the fate and response of cells to diverse signals. Therefore the identification of regulators which mediate these signals is a crucial step in elucidating how cell fate is regulated. We present here a detailed analysis of TPKs and BCY1 genes regulation based on the use of a dual reporter screen to discover regulators. In , three genes encode the PKA catalytic subunit, TPK1, TPK2, TPK3, while the regulatory subunit is encoded by only one gene, BCY1. Using a two-color GFP-RFP reporter system, Reporter-Synthetic Genetic Array, we systematically assesses the effects of gene deletions on the TPK1, TPK2, TPK3 and BCY1 promoters activity. We could define a list of positive and negative transcription regulators that were classified using Gene Ontology analysis into functionally related groups.We identified genes with roles in lipid and phosphate metabolism, cytoskeleton organization and transcription regulation. Genetic networks were constructed showing that several regulators are shared by the PKA subunits. The involvement of some of these pathways in TPKs and BCY1 transcription regulation was validated. Thus, the reporter gene assay provides a powerful means to link gene function to transcriptional control.