PKA-dependent regulation of Cdc25 RasGEF localization in budding yeast.
In Saccharomyces cerevisiae the Cdc25/Ras/cAMP pathway is involved in cell growth and proliferation regulation. Ras proteins are regulated by Ira1/2 GTPase activating proteins (GAPs) and Cdc25/Sdc25 guanine nucleotide exchange factors (GEFs). Most of cytosolic Cdc25 protein was found on internal membranes in exponentially growing cells, while upon incubation in a buffer with no nutrients it is re-localized to plasma membrane. The overexpression of Tpk1 PKA catalytic subunit also induces Cdc25 export from the nucleus, involving two serine residues near the Nuclear Localization Site (NLS): mutation of Ser(825) and Ser(826) to glutamate is sufficient to exclude physiologically expressed Cdc25 from the nucleus, mimicking Tpk1 overproduction effect. Mutation of these Ser residues to Ala abolishes the effect of nuclear export induced by Tpk1 overexpression on a Cdc25eGFP fusion. Moreover, mutation of these residues affects PKA-related phenotypes such as heat shock resistance, glycogen content and cell volume.
Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy.
Fiorella
F
Belotti
Renata
R
Tisi
Chiara
C
Paiardi
Silvia
S
Groppi
Enzo
E
Martegani
D021581:Active Transport, Cell Nucleus; D020134:Catalytic Domain; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D010766:Phosphorylation; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020666:ras-GRF1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
C542351:CDC25 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D020666:ras-GRF1; D017868:Cyclic AMP-Dependent Protein Kinases