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J. Biol. Chem., Vol. 276, Issue 27, 24531-24539, July 6, 2001
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and
From the Transcription Laboratory, Imperial Cancer Research Fund
Laboratories, 44 Lincoln's Inn Fields, London
WC2A 3PX, United Kingdom
Activation of the transcription factor serum
response factor (SRF) is dependent on Rho-controlled changes in
actin dynamics. We used pathway-specific inhibitors to compare the
roles of actin dynamics, extracellular signal-regulated kinase (ERK)
signaling, and phosphatidylinositol 3-kinase in signaling either
to SRF itself or to four cellular SRF target genes. Serum,
lysophosphatidic acid, platelet-derived growth factor, and phorbol
12-myristate 13-acetate (PMA) each activated transcription of a
stably integrated SRF reporter gene dependent on functional RhoA
GTPase. Inhibition of mitogen-activated protein kinase-ERK kinase
(MEK) signalling reduced activation of the SRF reporter by all stimuli
by about 50%, except for PMA, which was effectively blocked.
Inhibition of phosphatidylinositol 3-kinase slightly reduced reporter
activation by serum and lysophosphatidic acid but substantially
inhibited activation by platelet-derived growth factor and PMA.
Reporter induction by all stimuli was absolutely dependent on actin
dynamics. Regulation of the SRF (srf) and vinculin
(vcl) genes was similar to that of the SRF reporter gene;
activation by all stimuli was Rho-dependent and required
actin dynamics but was largely independent of MEK activity. In
contrast, activation of fos and egr1 occurred independently of RhoA and actin polymerization but was almost completely dependent on MEK activation. These results show that at
least two classes of SRF target genes can be distinguished on the basis
of their relative sensitivity to RhoA-actin and MEK-ERK signaling pathways.
Supported by a predoctoral award from the Overseas Research
Student Scheme of the United Kingdom Committee of Vice Chancellors and Principals.
§
To whom correspondence should be addressed: Transcription
Laboratory, Rm. 401, Imperial Cancer Research Fund Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, UK; Fax:
44-20-7269-3093.
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