Abstract
The importance of transforming growth factor-β1 (TGF-β1) in plasminogen activator inhibitor-1 (PAI-1) gene expression has been established, but the precise intracellular mechanisms are not fully understood. Our hypothesis is that the actin cytoskeleton is involved in TGF-β1/MAPK-mediated PAI-1 expression in human mesangial cells. Examination of the distributions of actin filaments (F-actin), α-actinin, extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) by immunofluorescence and immunoprecipitation revealed that ERK and JNK associate with α-actinin along F-actin and that TGF-β1 stimulation promote the dissociation of ERK and JNK with F-actin. Disassembly of the actin cytoskeleton inhibited phosphorylation of ERK and JNK and modulated PAI-1 expression and promoter activity under both basal and TGF-β1-stimulated conditions. Stabilizing actin prevented dephosphorylation of ERK and JNK. ERK and JNK inhibitors and overexpressed dominant negative mutants antagonized the ability of TGF-β1 to increase PAI-1 expression and promoter activity. Disassembly of F-actin also inhibited AP-1 DNA binding activity as determined by electrophoretic mobility shift assay using AP-1 consensus oligonucleotides derived from human PAI-1 promoter. F-actin stabilization prevented loss of AP-1 DNA binding activity. Therefore, changes in actin cytoskeleton modulate the ability of TGF-β1 to stimulate PAI-1 expression through a mechanism dependent on the activation of MAPK/AP-1 pathways.
Original language | English (US) |
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Pages (from-to) | 1240-1250 |
Number of pages | 11 |
Journal | Experimental Cell Research |
Volume | 313 |
Issue number | 6 |
DOIs | |
State | Published - Apr 1 2007 |
Externally published | Yes |
Keywords
- Actin cytoskeleton
- AP-1
- MAPK
- Plasminogen activator inhibitor-1
- Transforming growth factor-β1
- α-Actinin
ASJC Scopus subject areas
- Cell Biology