Coordinated increase in cellular RNA and protein content induced by overexpression of Far1, a cyclin dependent kinase inhibitor, involves large transcriptional reprogramming and requires the Sfp1 protein.

ABSTRACT: The FAR1 gene encodes a large protein, whose major function is inhibition of cyclin-dependent kinase complexes involved in the G1/S transition. It has been proposed that Far1, together with the G1 cyclin Cln3, may be part of a cell sizer mechanism that controls the entry into S phase. A genome-wide transcriptional analysis of FAR1-overexpressing and far1 deleted cells grown in ethanol- or glucose-supplemented minimal media indicates that FAR1 overexpression induces strong transcriptional remodelling, metabolism being the most affected cellular property. These data suggest that the Far1/Cln3 sizer regulates cell growth either directly or indirectly by affecting metabolism and pathways known to modulate ribosome biogenesis. A crucial role in mediating the effect of Far1 overexpression is played by the Sfp1 protein, a key transcriptional regulator of ribosome biogenesis, whose presence is mandatory to allow a coordinated increase in both RNA and protein levels in ethanol-grown cells. The experimental plan was designed to study the effect of alteration of dosage of the FAR1 gene on the transcriptional pattern of cells exponentially growing in media supplemented with either glucose or ethanol as a carbon source. To this end wild type cells (strain W303-1A), far1 deleted mutant (background W303-1A, relevant genotype far1::his3) and strain overexpressing FAR1 (background W303-1A, transformed with plasmid pTet-FAR1-15Myc (Alberghina et al., 2004)) were collected in independent triplicates in exponential growth as detailed in growth protocol. Microarray experiments were performed using a yeast S98 chip oligonucleotide array (Affymetrix, Inc.) according to the manufacturer's instructions (http://www.affymetrix.com/support/technical/manuals.affx).

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Marco Vanoni 

PROVIDER: E-GEOD-31143 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress