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A more recent version of this article appeared on March 12, 2004
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Papers In Press, published online ahead of print December 16, 2003
J. Biol. Chem, 10.1074/jbc.M313048200
Submitted on December 1, 2003
Revised on December 16, 2003
Accepted on December 16, 2003

VASP activation of SRE-dependent transcription occurs downstream of RhoA and is inhibited by cGMP-dependent protein kinase phosphorylation

Shunhui Zhuang, Giao T. Nguyen, Yongchang Chen, Tanima Gudi, Martin Eigenthaler, Thomas Jarchau, Ulrich Walter, Gerry R. Boss, and Renate B. Pilz

Medicine Dept., Univ. of California San Diego, La Jolla, CA 92093-0652

Corresponding Author: rpilz{at}ucsd.edu

Vasodilator-stimulated phosphoprotein (VASP) associates with cytoskeletal structures and promotes F-actin formation. RhoA, a member of the Ras superfamily of proteins, activates serum response element (SRE)-dependent transcription through changes in actin dynamics. We now show that the F-actin binding region of VASP is required for VASP stimulation of SRE-dependent transcription, and that VASP is downstream of RhoA in stimulating SRE-dependent transcription. The isolated carboxyl-terminal coiled-coil region of VASP mediates protein tetramerization and has been used as a dominant negative form of VASP; we found that it forms complexes with endogenous VASP in vivo and inhibits in a dose-dependent fashion serum-, RhoA- and VASP-stimulated SRE-dependent transcription. Cyclic GMP-dependent protein kinase (G-kinase) inhibits RhoA activation of SRE-dependent transcription (J.Biol.Chem.277:37382-93,2002). We now show that the G-kinase inhibition that occurs downstream of RhoA can be explained, at least in part, by G-kinase phosphorylation of VASP on Ser239 at the carboxyl-terminal end of the G-actin binding site, with some contribution by phosphorylation of Ser157, which is proximal to the profilin binding site. A phosphorylation-deficient VASP mutant can partly prevent cGMP/G-kinase inhibition of serum- and RhoA-induced SRE-dependent transcription. These studies show that VASP, an important component of the cellular microfilament system, plays a major role in regulating SRE-dependent transcription, and that G-kinase regulates VASP activity.


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