INVESTIGADORES
MORENO Silvia Margarita
artículos
Título:
Differential localization to cytoplasm, nucleus or P-bodies of yeast PKA subunits under different growth conditions.
Autor/es:
VANESA TUDISCA; VICTORIA RECOUVREUX; SILVIA MORENO; EMANUELLE BOY-MARCOTTE; MICHEL JACQUET; PAULA PORTELA,
Revista:
EUROPEAN JOURNAL OF CELL BIOLOGY
Editorial:
ELSEVIER GMBH
Referencias:
Año: 2010 vol. 89 p. 339 - 348
ISSN:
0171-9335
Resumen:
Our aiminthisworkwastofurthercharacterizethecomplexityandspecificityofthethreedifferent isoforms (Tpk1,Tpk2andTpk3)ofthecatalyticandregulatory(Bcy1)subunitsofPKAin Saccharomyces cerevisiae. WethusanalyzedthesubcellularlocalizationofthePKAsubunitsinlivingcellsbyusing strains carryingGFP(greenfluorescentprotein)fusedtoeachPKAsubunit.Duringexponentialgrowth on glucose,bothBcy1andTpk2localizedinthenucleus,whereasTpk1andTpk3showedamixed pattern ofnucleo-cytoplasmiclocalization.Duringexponentialgrowthonglycerolandduring stationaryphase,thePKAsubunitsshowedmostlycytoplasmiclocalization,withthepeculiaritythat Tpk2 andTpk3butnotBcy1,werefoundassociatedtoP-bodiesandEGPbodies.Tpk3wasaccumulated into P-bodiesduringglucosedeprivationandhyperosmoticstress.DeletionofTpk3alteredthekinetics of P-bodyformation.Analysisofproteinexpressionshowedthattherelativeexpressionpatternofeach Tpk changesfromlowlevelsunderfermentativemetabolismtohigherlevelsduringstationaryphase. The increaseinTpklevelsproducedanimbalancewithBcy1levels.Ourdatasuggestthatthesignaling specificitythroughPKAinyeastcouldbemediatedbyaparticularsubcellularlocalizationofeach isoform ofTpk. Saccharomyces cerevisiae. WethusanalyzedthesubcellularlocalizationofthePKAsubunitsinlivingcellsbyusing strains carryingGFP(greenfluorescentprotein)fusedtoeachPKAsubunit.Duringexponentialgrowth on glucose,bothBcy1andTpk2localizedinthenucleus,whereasTpk1andTpk3showedamixed pattern ofnucleo-cytoplasmiclocalization.Duringexponentialgrowthonglycerolandduring stationaryphase,thePKAsubunitsshowedmostlycytoplasmiclocalization,withthepeculiaritythat Tpk2 andTpk3butnotBcy1,werefoundassociatedtoP-bodiesandEGPbodies.Tpk3wasaccumulated into P-bodiesduringglucosedeprivationandhyperosmoticstress.DeletionofTpk3alteredthekinetics of P-bodyformation.Analysisofproteinexpressionshowedthattherelativeexpressionpatternofeach Tpk changesfromlowlevelsunderfermentativemetabolismtohigherlevelsduringstationaryphase. The increaseinTpklevelsproducedanimbalancewithBcy1levels.Ourdatasuggestthatthesignaling specificitythroughPKAinyeastcouldbemediatedbyaparticularsubcellularlocalizationofeach isoform ofTpk.