Ras Affects Filamentous Growth in Saccharomyces cerevisiae Solely Through Regulation of Protein Kinase A

ABSTRACT: To assess the relative contributions of the PKA and MAPK pathways in Ras stimulation of filamentation, we examined the change in transcriptional profile following activation of Ras in strains in which one of these two pathways has been severed. These arrays contain the experiments involving WT strain with glucose addition, Ras2G19V strain with galactose induction, tpk2-w strain as a control, and RasG19V activation in the tpk2-w background. Activation of Ras in the Σ1278b background – by galactose addition to a gal1 PGAL10-RAS2G19V strain - causes a massive restructuring of the transcription pattern of the cell, which closely resembles the changes induced by addition of glucose. To assess the extent to which these Ras-induced changes depended on signaling through PKA, we compared the expression profile following induction of activated Ras in a wild type background to that following induction of activated Ras in the bcy1 tpk1 tpk3 tpk2V218G. Keywords: time course RNA from strains grown as described was extracted using the RNeasy Mini Kit (Qiagen, California, USA) with mechanical disruption. RNA labeling and hybridization to Agilent oligonucleotide microarrays were done according to manufacturer’s instructions (Agilent Technologies, California, USA). RNA and cRNA concentrations were measured with NanoDrop (NanoDrop Technologies, Delaware, USA). 250ng labeled cRNA was used for each hybridization. The zero minute time point samples were labeled with Cy3 and others with Cy5. At least two technical replicates of each hybridization were performed. Hybridized microarrays were scanned with Agilent Microarray Scanner G2505B and the images were prepared for data analysis with Feature Extraction Software version 7.5 (Agilent Technologies, California, USA). Data were filtered and analyzed using PUMAdb based on the Stanford Microarray Database package.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Cemile Guldal 

PROVIDER: E-GEOD-8805 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress