title: Cytosolic stress reduces degradation of connexin43 internalized from the cell surface and enhances gap junction formation and function.
abstract: The protein constituents of gap junctions, connexins, have a rapid basal rate of degradation even after transport to the cell surface. We have used cell surface biotinylation to label gap junction-unassembled plasma membrane pools of connexin43 (Cx43) and show that their degradation is inhibited by mild hyperthermia, oxidative stress, and proteasome inhibitors. Cytosolic stress does not perturb endocytosis of biotinylated Cx43, but instead it seems to interfere with its targeting and/or transport to the lysosome, possibly by increasing the level of unfolded protein in the cytosol. This allows more Cx43 molecules to recycle to the cell surface, where they are assembled into long-lived, functional gap junctions in otherwise gap junction assembly-inefficient cells. Cytosolic stress also slowed degradation of biotinylated Cx43 in gap junction assembly-efficient normal rat kidney fibroblasts, and reduced the rate at which gap junctions disappeared from cell interfaces under conditions that blocked transport of nascent connexin molecules to the plasma membrane. These data demonstrate that degradation from the cell surface can be down-regulated by physiologically relevant forms of stress. For connexins, this may serve to enhance or preserve gap junction-mediated intercellular communication even under conditions in which protein synthesis and/or intracellular transport are compromised.
authors.affiliation: Division of Molecular Medicine, Oregon Health and Science University, Portland, OR 97239, USA.
authors.forename: Judy K
authors.initials: JK
authors.lastname: VanSlyke
authors.forename: Linda S
authors.initials: LS
authors.lastname: Musil
mesh_terms: D000818:Animals; D019921:Biotinylation; D016466:CHO Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D018031:Connexin 43 / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003600:Cytosol / Q000502:physiology; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism; D017629:Gap Junctions / Q000502:physiology*; D006358:Hot Temperature; D008247:Lysosomes / Q000378:metabolism / Q000502:physiology; D051379:Mice; D018384:Oxidative Stress / Q000502:physiology; D046988:Proteasome Endopeptidase Complex / Q000502:physiology; D017510:Protein Folding; D051381:Rats; D014162:Transfection; D015854:Up-Regulation
chemical_list: D018031:Connexin 43; D046988:Proteasome Endopeptidase Complex