Integrin dependence of brain natriuretic peptide gene promoter activation by mechanical strain.
Expression of the brain natriuretic peptide (BNP) gene in cultured neonatal rat ventricular myocytes is activated by mechanical strain in vitro. We explored the role of cell-matrix contacts in initiating the strain-dependent increment in human BNP (hBNP) promoter activity. Coating the culture surface with fibronectin effected a dose-dependent increase in basal hBNP luciferase activity and amplification of the response to strain. Preincubation of myocytes with an RGD peptide (GRGDSP) or with soluble fibronectin, each of which would be predicted to compete for cell-matrix interactions, resulted in a dose-dependent reduction in strain-dependent hBNP promoter activity. A functionally inert RGE peptide (GRGESP) was without effect. Using fluorescence-activated cell sorting, we demonstrated the presence of beta(1), beta(3), and alpha(v)beta(5) integrins in myocytes as well as non-myocytes and alpha1 only in non-myocytes in our cultures. Inclusion of antibodies directed against beta(1), beta(3), or alpha(v)beta(5), but not alpha(1), alpha(2), or cadherin, was effective in blocking the BNP promoter response to mechanical strain. These same antibodies (anti-beta(3), -beta(1), and -alpha(v)beta(5)) had a similar inhibitory effect on strain-stimulated ERK, p38 MAPK, and, to a lesser extent, JNK activities in these cells. Cotransfection with chimeric integrin receptors capable of acting as dominant-negative inhibitors of integrin function demonstrated suppression of strain-dependent BNP promoter activity when vectors encoding beta(1) or beta(3), but not beta(5), alpha(5), or a carboxyl-terminal deletion mutant of beta(3) (beta(3)B), were employed. These studies underscore the importance of cell-matrix interactions in controlling cardiac gene expression and suggest a potentially important role for these interactions in signaling responses to mechanical stimuli within the myocardium.
Liang
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Metabolic Research Unit, the Lung Biology Center, and the Department of Medicine, University of California, San Francisco, California 94143, USA.
Atakilit
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Gardner
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D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000494:pharmacology; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000235:genetics; D006352:Heart Ventricles / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000276:immunology / Q000378:metabolism*; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009154:Mutation; D020097:Natriuretic Peptide, Brain / Q000235:genetics*; D009842:Oligopeptides / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015398:Signal Transduction; D013314:Stress, Mechanical; D014162:Transfection
D000906:Antibodies; D005353:Fibronectins; D016023:Integrins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C112658:arginyl-glycyl-glutamic acid; D020097:Natriuretic Peptide, Brain; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases