Journal
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
Volume 1833, Issue 12, Pages 3254-3264Publisher
ELSEVIER
DOI: 10.1016/j.bbamcr.2013.09.014
Keywords
Cell cycle; RNA Polymerase II; Saccharomyces cerevisiae; SBF-MBF complexes; Snf1/AMPK; SNF1(as) (I132G)
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Funding
- EU
- MIUR
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Snf1, the yeast AMP-activated kinase homolog, regulates the expression of several genes involved in adaptation to glucose limitation and in response to cellular stresses. We previously demonstrated that Snf1 interacts with Swi6, the regulatory subunit of SBF and MBF complexes, and activates CLB5 transcription. Here we report that, in alpha-factor synchronized cells in 2% glucose, the loss of the Snfl catalytic subunit impairs the binding of SBF and MBF complexes and the subsequent recruitment of the FACT complex and RNA Polymerase II to promoters of G1-genes. By using an analog-sensitive allele of SNF1, SNF1(as)(I132G), encoding a protein whose catalytic activity is selectively inhibited in vivo by 2-naphthylmethyl pyrazolopyrimidine 1, we show that the inhibition of Snfl catalytic activity affects the expression of G1-genes causing a delayed entrance into S phase in cells synchronized in G1 phase by a-factor treatment or by elutriation. Moreover, Snfl is detected in immune complexes of Rpb1, the large subunit of RNA Polymerase II, and is present at both promoters and coding regions of SBF- and MBF-regulated genes 20 min after alpha-factor release, suggesting a direct role for Snfl in the activation of the Glregulon transcription. (C) 2013 Elsevier B.V. All rights reserved.
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