Regulation of cytoskeleton organization and paxillin dephosphorylation by cAMP. Studies on murine Y1 adrenal cells.
Cyclic AMP induces corticosteroid production, differential gene transcription, and cell cycle arrest in adrenal cortex-derived Y1 cells. These responses follow a cAMP-controlled transformation in Y1 cell morphology: the conversion of flat epithelial cells into rounded, highly refractile cells with short processes. Little is known about effector proteins and mechanisms that link activated protein kinase A to the alteration in cell shape. We now report that cAMP causes rapid (</=1 min) and selective tyrosine dephosphorylation of paxillin, a focal adhesion protein. Paxillin is maximally dephosphorylated before other physiological effects of cAMP are detected in Y1 cells. Dephosphopaxillin translocates from focal adhesions to the cytoplasm as stress fibers vanish and F-actin accumulates in membrane ruffles and cytoplasmic aggregates. Remnants of focal adhesion complexes dissociate from the cell cortex and coalesce into large structures that contain aggregated F-actin. Pervanadate, an inhibitor of protein-tyrosine phosphatases, abrogates all effects of cAMP. Conversely, genistein-sensitive protein-tyrosine kinase activity is essential for establishing epithelial morphology and reversing effects of cAMP in Y1 cells. Thus, cAMP/protein kinase A (PKA) actions are initially targeted to focal adhesions and cortical actin cytoskeleton; paxillin is an early and unexpected downstream target in a PKA-mediated signaling pathway, and protein-tyrosine phosphatase activity provides an essential link between PKA activation and the control of cell shape.
Department of Molecular Pharmacology, Atran Laboratories, Albert Einstein College of Medicine, Bronx, New York 10461, USA. rubin@aecom.yu.edu
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D000199:Actins / Q000378:metabolism; D000306:Adrenal Cortex Neoplasms / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism*; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019833:Genistein; D007529:Isoflavones / Q000494:pharmacology; D051379:Mice; D051419:Paxillin; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000276:immunology / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism; D014638:Vanadates / Q000494:pharmacology
D000199:Actins; D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; D007529:Isoflavones; D051419:Paxillin; D010750:Phosphoproteins; C496062:Pxn protein, mouse; C076715:pervanadate; D014638:Vanadates; D014443:Tyrosine; D019833:Genistein; D000242:Cyclic AMP; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse; D017868:Cyclic AMP-Dependent Protein Kinases