Loss of functional caveolae during senescence of human fibroblasts.
Primary human fibroblasts have a finite replicative lifespan in culture that culminates in a unique state of growth arrest, termed senescence that is accompanied by distinct morphological and biochemical alterations. Senescent cell responses to extracellular stimuli are believed to be altered at a point after receptors are bound by ligand, leading to improper integration of the signals which initiate DNA replication. In this study we demonstrate that one of the key organizing membrane microdomains for receptor signaling, caveolae, are absent in senescent cells. A comparison of young and senescent cells indicated that senescent cells contained a higher total amount of caveolins 1 and 2 but had significantly less of both proteins in the caveolar fraction. Additionally, caveolar fractions from senescent cells completely lacked the tyrosine-kinase activity associated with functional caveolae. Furthermore, old cells had little caveolar protein exposed to the outer plasma membrane as estimated by using an in vivo biotinylation assay and no detectable caveolin 1 on the cell surface when processed for immunofluoresence and confocal microscopy. Together, these data suggest that a fundamental loss of signal integration at the plasma membrane of senescent cells is due to the loss of signaling competent caveolae.
Department of Biochemistry and Molecular Biology, Southern Alberta Cancer Centre, University of Calgary, Heritage Medical Research Building, Calgary Alberta, Canada.
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Wheaton
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Sampsel
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Boisvert
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Davy
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Robbins
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Riabowol
D021941:Caveolae / Q000737:chemistry / Q000201:enzymology*; D051242:Caveolin 1; D051243:Caveolin 2; D022461:Caveolins / Q000032:analysis / Q000378:metabolism; D002458:Cell Fractionation; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology*; D003902:Detergents; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005347:Fibroblasts / Q000648:ultrastructure; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry / Q000201:enzymology; D017830:Octoxynol; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051076:Proto-Oncogene Proteins c-fyn; D013395:Sucrose
C495540:CAV1 protein, human; D051242:Caveolin 1; D051243:Caveolin 2; D022461:Caveolins; D003902:Detergents; D011518:Proto-Oncogene Proteins; D013395:Sucrose; D004815:Epidermal Growth Factor; D017830:Octoxynol; D011505:Protein-Tyrosine Kinases; C494957:FYN protein, human; D051076:Proto-Oncogene Proteins c-fyn