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Published 20 August 2001. doi:10.1083/jcb.200103011
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© The Rockefeller University Press, 0021-9525/2001/8/753 $5.00
The Journal of Cell Biology, Volume 154, Number 4, August 20, 2001 753-762

Article

Disruption of cytoskeletal integrity impairs Gi-mediated signaling due to displacement of Gi proteins



Wilhelm Bloch1, Yun Fan2, Ji Han1, Sheng Xue2, Torsten Schöneberg3, Guanju Ji2, Zhong J. Lu2, Michael Walther1, Reinhard Fässler4, Jürgen Hescheler2, Klaus Addicks1 and Bernd K. Fleischmann2

1 Institute of Anatomy I, University of Cologne, Germany
2 Institute of Neurophysiology, University of Cologne, Germany
3 Institute of Pharmacology, Free University of Berlin, Germany
4 Department of Experimental Pathology, University of Lund, Sweden

Address correspondence to B.K. Fleischmann, Institute of Neurophysiology, University of Cologne, Robert-Kochstr. 39, D-50931 Cologne, Germany. Tel.: 49-221-478-6940. Fax: 49-221-478-3834. E-mail: akp17{at}uni-koeln.de

ß1 integrins play a crucial role as cytoskeletal anchorage proteins. In this study, the coupling of the cytoskeleton and intracellular signaling pathways was investigated in ß1 integrin deficient (-/-) embryonic stem cells. Muscarinic inhibition of the L-type Ca2+ current (ICa) and activation of the acetylcholine-activated K+ current (IK,ACh) was found to be absent in ß1 integrin-/- cardiomyocytes. Conversely, ß adrenoceptor-mediated modulation of ICa was unaffected by the absence of ß1 integrins. This defect in muscarinic signaling was due to defective G protein coupling. This was supported by deconvolution microscopy, which demonstrated that Gi exhibited an atypical subcellular distribution in the ß1 integrin-/- cardiomyocytes. A critical role of the cytoskeleton was further demonstrated using cytochalasin D, which displaced Gi and impaired muscarinic signaling. We conclude that cytoskeletal integrity is required for correct localization and function of Gi-associated signaling microdomains.

Key Words: embryonic stem cells; signal transduction; ion channels; G proteins; ß1 integrins


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