Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly

Joanne E. Murphy-Ullrich, Manuel A. Pallero, Nancy Boerth, Jeffrey A. Greenwood, Thomas M. Lincoln, Trudy L. Cornwell

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Focal adhesions are specialized regions of cell membranes that are foci for the transmission of signals between the outside and the inside of the cell. Intracellular signaling events are important in the organization and stability of these structures. In previous work, we showed that the counter-adhesive extracellular matrix proteins, thrombospondin, tenascin, and SPARC, induce the disassembly of focal adhesion plaques and we identified the active regions of these proteins. In order to determine the mechanisms whereby the anti-adhesive matrix proteins modulate cytoskeletal organization and focal adhesion integrity, we examined the role of protein kinases in mediating the loss of focal adhesions by these proteins. Data from these studies show that cGMP-dependent protein kinase is necessary to mediate focal adhesion disassembly triggered by either thrombospondin or tenascin, but not by SPARC. In experiments using various protein kinase inhibitors, we observed that selective inhibitors of cyclic GMP-dependent protein kinase, KT5823 and Rp-8-Br-cGMPS, blocked the effects of both the active sequence of thrombospondin 1 (hep I) and the alternatively-spliced segment (TNfA-D) of tenascin-C on focal adhesion disassembly. Moreover, early passage rat aortic smooth muscle cells which have high levels of cGMP-dependent protein kinase were sensitive to hep I treatment, in contrast to passaged cGMP-dependent protein kinase deficient cells which were refractory to hep I or TNfnA-D treatment, but were sensitive to SPARC. Transfection of passaged smooth muscle cells with the catalytic domain of PKG Iα restored responsiveness to hep I and TNfnA-D. While these studies show that cGMP-dependent protein kinase activity is necessary for thrombospondin and tenascin-mediated focal adhesion disassembly, kinase activity alone is not sufficient to induce disassembly as transfection of the catalytic domain of the kinase in the absence of additional stimuli does not result in loss of focal adhesions.

Original languageEnglish (US)
Pages (from-to)2499-2508
Number of pages10
JournalJournal of Cell Science
Volume109
Issue number10
StatePublished - Oct 1 1996
Externally publishedYes

Fingerprint

Thrombospondins
Tenascin
Cyclic GMP-Dependent Protein Kinases
Focal Adhesions
Adhesives
Smooth Muscle Myocytes
Transfection
Catalytic Domain
Thrombospondin 1
Dilatation and Curettage
Focal Adhesion Protein-Tyrosine Kinases
Cytoskeletal Proteins
Extracellular Matrix Proteins
Protein Kinase Inhibitors
Protein Kinases
Proteins
Phosphotransferases
Cell Membrane

Keywords

  • Focal adhesion
  • PKG
  • SPARC
  • Tenascin
  • Thrombospondin

ASJC Scopus subject areas

  • Cell Biology

Cite this

Murphy-Ullrich, J. E., Pallero, M. A., Boerth, N., Greenwood, J. A., Lincoln, T. M., & Cornwell, T. L. (1996). Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly. Journal of Cell Science, 109(10), 2499-2508.

Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly. / Murphy-Ullrich, Joanne E.; Pallero, Manuel A.; Boerth, Nancy; Greenwood, Jeffrey A.; Lincoln, Thomas M.; Cornwell, Trudy L.

In: Journal of Cell Science, Vol. 109, No. 10, 01.10.1996, p. 2499-2508.

Research output: Contribution to journalArticle

Murphy-Ullrich, JE, Pallero, MA, Boerth, N, Greenwood, JA, Lincoln, TM & Cornwell, TL 1996, 'Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly', Journal of Cell Science, vol. 109, no. 10, pp. 2499-2508.
Murphy-Ullrich, Joanne E. ; Pallero, Manuel A. ; Boerth, Nancy ; Greenwood, Jeffrey A. ; Lincoln, Thomas M. ; Cornwell, Trudy L. / Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly. In: Journal of Cell Science. 1996 ; Vol. 109, No. 10. pp. 2499-2508.
@article{23cd886d89fa4a15b9203583da04a4d0,
title = "Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly",
abstract = "Focal adhesions are specialized regions of cell membranes that are foci for the transmission of signals between the outside and the inside of the cell. Intracellular signaling events are important in the organization and stability of these structures. In previous work, we showed that the counter-adhesive extracellular matrix proteins, thrombospondin, tenascin, and SPARC, induce the disassembly of focal adhesion plaques and we identified the active regions of these proteins. In order to determine the mechanisms whereby the anti-adhesive matrix proteins modulate cytoskeletal organization and focal adhesion integrity, we examined the role of protein kinases in mediating the loss of focal adhesions by these proteins. Data from these studies show that cGMP-dependent protein kinase is necessary to mediate focal adhesion disassembly triggered by either thrombospondin or tenascin, but not by SPARC. In experiments using various protein kinase inhibitors, we observed that selective inhibitors of cyclic GMP-dependent protein kinase, KT5823 and Rp-8-Br-cGMPS, blocked the effects of both the active sequence of thrombospondin 1 (hep I) and the alternatively-spliced segment (TNfA-D) of tenascin-C on focal adhesion disassembly. Moreover, early passage rat aortic smooth muscle cells which have high levels of cGMP-dependent protein kinase were sensitive to hep I treatment, in contrast to passaged cGMP-dependent protein kinase deficient cells which were refractory to hep I or TNfnA-D treatment, but were sensitive to SPARC. Transfection of passaged smooth muscle cells with the catalytic domain of PKG Iα restored responsiveness to hep I and TNfnA-D. While these studies show that cGMP-dependent protein kinase activity is necessary for thrombospondin and tenascin-mediated focal adhesion disassembly, kinase activity alone is not sufficient to induce disassembly as transfection of the catalytic domain of the kinase in the absence of additional stimuli does not result in loss of focal adhesions.",
keywords = "Focal adhesion, PKG, SPARC, Tenascin, Thrombospondin",
author = "Murphy-Ullrich, {Joanne E.} and Pallero, {Manuel A.} and Nancy Boerth and Greenwood, {Jeffrey A.} and Lincoln, {Thomas M.} and Cornwell, {Trudy L.}",
year = "1996",
month = "10",
day = "1",
language = "English (US)",
volume = "109",
pages = "2499--2508",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "10",

}

TY - JOUR

T1 - Cyclic GMP-dependent protein kinase is required for thrombospondin and tenascin mediated focal adhesion disassembly

AU - Murphy-Ullrich, Joanne E.

AU - Pallero, Manuel A.

AU - Boerth, Nancy

AU - Greenwood, Jeffrey A.

AU - Lincoln, Thomas M.

AU - Cornwell, Trudy L.

PY - 1996/10/1

Y1 - 1996/10/1

N2 - Focal adhesions are specialized regions of cell membranes that are foci for the transmission of signals between the outside and the inside of the cell. Intracellular signaling events are important in the organization and stability of these structures. In previous work, we showed that the counter-adhesive extracellular matrix proteins, thrombospondin, tenascin, and SPARC, induce the disassembly of focal adhesion plaques and we identified the active regions of these proteins. In order to determine the mechanisms whereby the anti-adhesive matrix proteins modulate cytoskeletal organization and focal adhesion integrity, we examined the role of protein kinases in mediating the loss of focal adhesions by these proteins. Data from these studies show that cGMP-dependent protein kinase is necessary to mediate focal adhesion disassembly triggered by either thrombospondin or tenascin, but not by SPARC. In experiments using various protein kinase inhibitors, we observed that selective inhibitors of cyclic GMP-dependent protein kinase, KT5823 and Rp-8-Br-cGMPS, blocked the effects of both the active sequence of thrombospondin 1 (hep I) and the alternatively-spliced segment (TNfA-D) of tenascin-C on focal adhesion disassembly. Moreover, early passage rat aortic smooth muscle cells which have high levels of cGMP-dependent protein kinase were sensitive to hep I treatment, in contrast to passaged cGMP-dependent protein kinase deficient cells which were refractory to hep I or TNfnA-D treatment, but were sensitive to SPARC. Transfection of passaged smooth muscle cells with the catalytic domain of PKG Iα restored responsiveness to hep I and TNfnA-D. While these studies show that cGMP-dependent protein kinase activity is necessary for thrombospondin and tenascin-mediated focal adhesion disassembly, kinase activity alone is not sufficient to induce disassembly as transfection of the catalytic domain of the kinase in the absence of additional stimuli does not result in loss of focal adhesions.

AB - Focal adhesions are specialized regions of cell membranes that are foci for the transmission of signals between the outside and the inside of the cell. Intracellular signaling events are important in the organization and stability of these structures. In previous work, we showed that the counter-adhesive extracellular matrix proteins, thrombospondin, tenascin, and SPARC, induce the disassembly of focal adhesion plaques and we identified the active regions of these proteins. In order to determine the mechanisms whereby the anti-adhesive matrix proteins modulate cytoskeletal organization and focal adhesion integrity, we examined the role of protein kinases in mediating the loss of focal adhesions by these proteins. Data from these studies show that cGMP-dependent protein kinase is necessary to mediate focal adhesion disassembly triggered by either thrombospondin or tenascin, but not by SPARC. In experiments using various protein kinase inhibitors, we observed that selective inhibitors of cyclic GMP-dependent protein kinase, KT5823 and Rp-8-Br-cGMPS, blocked the effects of both the active sequence of thrombospondin 1 (hep I) and the alternatively-spliced segment (TNfA-D) of tenascin-C on focal adhesion disassembly. Moreover, early passage rat aortic smooth muscle cells which have high levels of cGMP-dependent protein kinase were sensitive to hep I treatment, in contrast to passaged cGMP-dependent protein kinase deficient cells which were refractory to hep I or TNfnA-D treatment, but were sensitive to SPARC. Transfection of passaged smooth muscle cells with the catalytic domain of PKG Iα restored responsiveness to hep I and TNfnA-D. While these studies show that cGMP-dependent protein kinase activity is necessary for thrombospondin and tenascin-mediated focal adhesion disassembly, kinase activity alone is not sufficient to induce disassembly as transfection of the catalytic domain of the kinase in the absence of additional stimuli does not result in loss of focal adhesions.

KW - Focal adhesion

KW - PKG

KW - SPARC

KW - Tenascin

KW - Thrombospondin

UR - http://www.scopus.com/inward/record.url?scp=0029909499&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029909499&partnerID=8YFLogxK

M3 - Article

C2 - 8923211

AN - SCOPUS:0029909499

VL - 109

SP - 2499

EP - 2508

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 10

ER -