Proteasome involvement in the degradation of the G(q) family of Galpha subunits.
Metabolically unstable proteins are involved in a multitude of regulatory networks, including those that control cell signaling, the cell cycle and in many responses to physiological stress. In the present study, we have determined the stability and characterized the degradation process of some members of the G(q) class of heterotrimeric G proteins. Pulse-chase experiments in HEK293 cells indicated a rapid turnover of endogenously expressed Galpha(q) and overexpressed Galpha(q) and Galpha(16) subunits. Pretreatment with proteasome inhibitors attenuated the degradation of both G alpha subunits. In contrast, pretreatment of cells with inhibitors of lysosomal proteases and nonproteasomal cysteine proteases had very little effect on the stability of the proteins. Significantly, the turnover of these proteins is not affected by transient activation of their associated receptors. Fractionation studies showed that the rates of Galpha(q) and Galpha16 degradation are accelerated in the cytosol. In fact, we show that a mutant Galpha(q) which lacks its palmitoyl modification site, and which is localized almost entirely in the cytoplasm, has a marked increase in the rate of degradation. Taken together, these results suggest that the G(q) class proteins are degraded through the proteasome pathway and that cellular localization and/or other protein interactions determine their stability.
Johansson
Bente B
BB
Department of Biomedicine, Faculty of Medicine, University of Bergen, Norway.
Minsaas
Laura
L
Aragay
Anna M
AM
D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D044385:GTP-Binding Protein alpha Subunits / Q000235:genetics / Q000378:metabolism*; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D043643:Receptor, Muscarinic M1 / Q000235:genetics / Q000378:metabolism; D043585:Receptor, Muscarinic M2 / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D015853:Cysteine Proteinase Inhibitors; D044385:GTP-Binding Protein alpha Subunits; D007295:Inositol Phosphates; D061988:Proteasome Inhibitors; D043643:Receptor, Muscarinic M1; D043585:Receptor, Muscarinic M2; D002217:Carbachol; D046988:Proteasome Endopeptidase Complex; D043802:GTP-Binding Protein alpha Subunits, Gq-G11