TY - JOUR
T1 - Snf1/AMPK promotes SBF and MBF-dependent transcription in budding yeast
AU - Busnelli, Sara
AU - Tripodi, Farida
AU - Nicastro, Raffaele
AU - Cirulli, Claudia
AU - Tedeschi, Gabriella
AU - Pagliarin, Roberto
AU - Alberghina, Lilia
AU - Coccetti, Paola
PY - 2013/12
Y1 - 2013/12
N2 - 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 α-factor synchronized cells in 2% glucose, the loss of the Snf1 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, SNF1as(I132G), encoding a protein whose catalytic activity is selectively inhibited in vivo by 2-naphthylmethyl pyrazolopyrimidine 1, we show that the inhibition of Snf1 catalytic activity affects the expression of G1-genes causing a delayed entrance into S phase in cells synchronized in G1 phase by α-factor treatment or by elutriation. Moreover, Snf1 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 α-factor release, suggesting a direct role for Snf1 in the activation of the G1-regulon transcription.
AB - 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 α-factor synchronized cells in 2% glucose, the loss of the Snf1 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, SNF1as(I132G), encoding a protein whose catalytic activity is selectively inhibited in vivo by 2-naphthylmethyl pyrazolopyrimidine 1, we show that the inhibition of Snf1 catalytic activity affects the expression of G1-genes causing a delayed entrance into S phase in cells synchronized in G1 phase by α-factor treatment or by elutriation. Moreover, Snf1 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 α-factor release, suggesting a direct role for Snf1 in the activation of the G1-regulon transcription.
KW - Cell cycle
KW - RNA Polymerase II
KW - Saccharomyces cerevisiae
KW - SBF-MBF complexes
KW - Snf1/AMPK
KW - SNF1(I132G)
UR - http://www.scopus.com/inward/record.url?scp=84885781874&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84885781874&partnerID=8YFLogxK
U2 - 10.1016/j.bbamcr.2013.09.014
DO - 10.1016/j.bbamcr.2013.09.014
M3 - Article
C2 - 24084603
AN - SCOPUS:84885781874
VL - 1833
SP - 3254
EP - 3264
JO - Biochimica et Biophysica Acta - Molecular Cell Research
JF - Biochimica et Biophysica Acta - Molecular Cell Research
SN - 0167-4889
IS - 12
ER -