Regulation of mTORC1 Signaling by pH 英文参考文献.docVIP

RegulationofmTORC1SignalingbypH ArunaD.Balgi1,GrahamH.Diering1,ElizabethDonohue1,KarenK.Y.Lam1,BrunoD.Fonseca1¤ ,Carla Zimmerman1,MasayukiNumata1,MichelRoberge1,2* 1Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada, 2The Centre for Drug Research and Development,Vancouver,BritishColumbia,Canada Abstract Background:Acidificationofthecytoplasmandtheextracellularenvironmentisassociatedwithmanyphysiologicaland pathological conditions, such as intense exercise, hypoxia and tumourigenesis. Acidification affects important cellular functionsincludingproteinsynthesis,growth,andproliferation.ManyofthesevitalfunctionsarecontrolledbymTORC1,a master regulator protein kinase that is activated by various growth-stimulating signals and inactivated by starvation conditions. Whether mTORC1 can also respond to changes in extracellular or cytoplasmic pH and play a role in limiting anabolicprocessesinacidicconditionsisnotknown. Methodology/Findings: We examined the effects of acidifying the extracellular medium from pH7.4 to 6.4 on human breast carcinoma MCF-7 cells and immortalized mouse embryo fibroblasts. Decreasing the extracellular pH caused intracellular acidification and rapid, graded and reversible inhibition of mTORC1, assessed by measuring the phosphorylationofthemTORC1substrateS6K.FibroblastsdeletedofthetuberoussclerosiscomplexTSC2gene,amajor negativeregulatorofmTORC1,wereunabletoinhibitmTORC1inacidicextracellularconditions,showingthattheTSC1– TSC2complexisrequiredforthisresponse.ExaminationofthemajorupstreampathwaysconvergingontheTSC1–TSC2 complexshowedthatAktsignalingwasunaffectedbypHbutthattheRaf/MEK/ERKpathwaywasinhibited.Inhibitionof MEKwithdrugscausedonlymodestmTORC1inhibition,implyingthatotherunidentifiedpathwaysalsoplaymajorroles. Conclusions:ThisstudyrevealsanovelrolefortheTSC1/TSC2complexandmTORC1insensingvariationsinambientpH. Asacommonfeatureoflowtissueperfusion,lowglucoseavailabilityandhighen