Lamb, N. J., Fernandez, A., Conti, M. A., Adelstein, R., Glass, D. B., Welch, W. J., Feramisco, J. R. (June 1988) Regulation of actin microfilament integrity in living nonmuscle cells by the cAMP-dependent protein kinase and the myosin light chain kinase. J Cell Biol, 106 (6). pp. 1955-71. ISSN 0021-9525
Abstract
Microinjection of the catalytic subunit of cAMP-dependent protein kinase (A-kinase) into living fibroblasts or the treatment of these cells with agents that elevate the intracellular cAMP level caused marked alterations in cell morphology including a rounded phenotype and a complete loss of actin microfilament bundles. These effects were transient and fully reversible. Two-dimensional gel electrophoresis was used to analyze the changes in phosphoproteins from cells injected with A-kinase. These experiments showed that accompanying the disassembly of actin microfilaments, phosphorylation of myosin light chain kinase (MLCK) increased and concomitantly, the phosphorylation of myosin P-light chain decreased. Moreover, inhibiting MLCK activity via microinjection of affinity-purified antibodies specific to native MLCK caused a complete loss of microfilament bundle integrity and a decrease in myosin P-light chain phosphorylation, similar to that seen after injection of A-kinase. These data support the idea that A-kinase may regulate microfilament integrity through the phosphorylation and inhibition of MLCK activity in nonmuscle cells.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Actins/*physiology Animals Cytoskeleton/*physiology Enzyme Activation Fluorescent Antibody Technique Immunologic Techniques In Vitro Microfilaments/*physiology Microinjections Microscopy, Electron Myosin-Light-Chain Kinase/*physiology Myosins/*physiology Phosphoproteins/physiology Protein Kinases/*physiology Rats Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Time Factors |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > actin Investigative techniques and equipment > assays > gel electrophoresis bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > kinase bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > myosin |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Gail Sherman |
| Date: | June 1988 |
| Date Deposited: | 28 Sep 2017 19:08 |
| Last Modified: | 28 Sep 2017 19:08 |
| PMCID: | PMC2115126 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/35172 |
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