Multiple TORC1-Associated Proteins Regulate Nitrogen Starvation-Dependent Cellular Differentiation in Saccharomyces cerevisiae 英文参考文献.docVIP

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Multiple TORC1-Associated Proteins Regulate Nitrogen Starvation-Dependent Cellular Differentiation in Saccharomyces cerevisiae 英文参考文献.doc

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Multiple TORC1-Associated Proteins Regulate Nitrogen Starvation-Dependent Cellular Differentiation in Saccharomyces cerevisiae 英文参考文献

MultipleTORC1-AssociatedProteinsRegulateNitrogen Starvation-DependentCellularDifferentiationin Saccharomycescerevisiae SunilLaxman*,BenjaminP.Tu* DepartmentofBiochemistry,UniversityofTexasSouthwesternMedicalCenter,Dallas,Texas,UnitedStatesofAmerica Abstract Background:ThebuddingyeastSaccharomycescerevisiaeundergoesdifferentiationintofilamentous-likeformsandinvades the growth medium as aforaging response tonutrient and environmental stresses. These developmental responses are under the downstream control of effectors regulated by the cAMP/PKA and MAPK pathways. However, the upstream sensors and signals that induce filamentous growth through these signaling pathways are not fully understood. Herein, through a biochemical purification of the yeast TORC1 (Target of Rapamycin Complex 1), we identify several proteins implicated in yeast filamentous growth that directly associate with the TORC1 and investigate their roles in nitrogen starvation-dependentorindependentdifferentiationinyeast. Methodology: We isolated the endogenous TORC1 by purifying tagged, endogenous Kog1p, and identified associated proteinsbymassspectrometry.WeestablishedinvasiveandpseudohyphalgrowthconditionsintwoS.cerevisiaegenetic backgrounds(S1278bandCEN.PK).Usingwildtypeandmutantstrainsfromthesegeneticbackgrounds,weinvestigated the roles of TORC1 and associated proteins in nitrogen starvation-dependent diploid pseudohyphal growth as well as nitrogenstarvation-independenthaploidinvasivegrowth. Conclusions:WeshowthatseveralproteinsidentifiedasassociatedwiththeTORC1areimportantfornitrogenstarvation- dependentdiploidpseudohyphalgrowth.Incontrast,invasivegrowthduetoothernutritionalstresseswasgenerallynot affectedinmutantstrainsoftheseTORC1-associatedproteins.OurstudiessuggestaroleforTORC1inyeastdifferentiation uponnitrogenstarvation.OurstudiesalsosuggesttheCEN.PKstrainbackgroundofS.cerevisiaemaybeparticularlyuseful forinvestigationsofnitrogenstarvation-induceddiploidpseudohyphalgrowth. Citation: Laxman S