This Article Full Text Full Text (PDF) Data Supplement All Versions of this Article: 26/7/1488    most recent 01.ATV.0000219615.88323.b4v1 Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Hasegawa, K. Articles by Hayashi, K. Search for Related Content PubMed PubMed Citation Articles by Hasegawa, K. Articles by Hayashi, K. Pubmed/NCBI databases Substance via MeSH Related Collections Angiogenesis Endothelium/vascular type/nitric oxide Related Article

(Arteriosclerosis, Thrombosis, and Vascular Biology. 2006;26:1488.)
© 2006 American Heart Association, Inc.

Vascular Biology

Dimethylarginine Dimethylaminohydrolase 2 Increases Vascular Endothelial Growth Factor Expression Through Sp1 Transcription Factor in Endothelial Cells

Kazuhiro Hasegawa; Shu Wakino; Toru Tanaka; Masumi Kimoto; Satoru Tatematsu; Takeshi Kanda; Kyoko Yoshioka; Koichiro Homma; Naoki Sugano; Masahiko Kurabayashi; Takao Saruta; Koichi Hayashi

From the Department of Internal Medicine (K. Hasegawa, S.W., S.T., T.K., K.Y., K. Homma, N.S., T.S., K. Hayashi), Keio University, Tokyo, Japan; Department of Medicine and Biological Science (T.T., M. Kurabayashi), Gunma University Graduate School of Medicine, Japan; and Department of Nutritional Science (M. Kimoto), Okayama Prefectural University, Japan.

Correspondence to Koichi Hayashi, MD, Department of Internal Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo, Japan. E-mail khayashi{at}sc.itc.keio.ac.jp

Objectives— Dimethylarginie dimethylaminohydrolase (DDAH) is a degrading enzyme for asymmetrical dimethylarginine, an endogenous NO synthase inhibitor. The molecular mechanism for DDAH-induced vascular endothelial growth factor (VEGF) expression was examined.

Methods and Results— Although the transfection of expression vectors for 2 isoforms of DDAH, DDAH1, or DDAH2 increased DDAH activity in bovine aortic endothelial cells and human umbilical vein endothelial cells, expression and secretion of VEGF were increased only in DDAH2-transfected cells. Knocking down the DDAH2 gene reduced VEGF production, and DDAH2 overexpression enhanced both proliferation and migration of endothelial cells. The VEGF promoter activity was increased by DDAH2 transfection, which was not blocked by an NO synthase (NOS) inhibitor but required the Sp1 sites. DDAH2 overexpression increased nuclear protein levels bound to Sp1 oligonucleotides in endothelial cells. Sp1 small interfering RNA blocked DDAH2-induced upregulation of VEGF. DDAH2 transfection increased nuclear and threonine-phosphorylation levels of Sp1 in a protein kinase A (PKA)–dependent manner. Protein–protein interaction between DDAH2 and PKA was enhanced in DDAH2-transfected cells.

Conclusions— DDAH2 upregulated the expression of VEGF through Sp1-dependent and NO/NOS system-independent promoter activation. DDAH2-increased Sp1 DNA binding activity was PKA dependent. These mechanisms may provide a novel therapeutic strategy for VEGF-related vasculopathies such as atherosclerosis.

This study demonstrates PKA- and Sp1-dependent transcriptional upregulation of VEGF by DDAH2 in vascular endothelial cells. This mechanism is specific for DDAH2 and independent of NO/NOS system. The induction of VEGF by DDAH2 may contribute to DDAH-induced angiogenesis and constitute a novel therapeutic target of angiogenesis-related diseases.

Key Words: DDAH • ADMA • Sp1 • VEGF • endothelial cell

Related Article:

New Tricks From an Old Dog: Nitric Oxide-Independent Effects of Dimethylarginine Dimethylaminohydrolase

James Leiper and Patrick Vallance
Arterioscler Thromb Vasc Biol 2006 26: 1419-1420. [Extract] [Full Text] [PDF]

This article has been cited by other articles:

				L. R Fiedler, T. Bachetti, J. Leiper, I. Zachary, L. Chen, T. Renne, and B. Wojciak-Stothard The ADMA/DDAH Pathway Regulates VEGF-Mediated Angiogenesis Arterioscler Thromb Vasc Biol, December 1, 2009; 29(12): 2117 - 2124. [Abstract] [Full Text] [PDF]

				M. R. Kim, H. S. Choi, J. W. Yang, B. C. Park, J.-A. Kim, and K. W. Kang Enhancement of vascular endothelial growth factor-mediated angiogenesis in tamoxifen-resistant breast cancer cells: role of Pin1 overexpression Mol. Cancer Ther., August 1, 2009; 8(8): 2163 - 2171. [Abstract] [Full Text] [PDF]

				F. Palm, M. L. Onozato, Z. Luo, and C. S. Wilcox Dimethylarginine dimethylaminohydrolase (DDAH): expression, regulation, and function in the cardiovascular and renal systems Am J Physiol Heart Circ Physiol, December 1, 2007; 293(6): H3227 - H3245. [Abstract] [Full Text] [PDF]

				K. Hasegawa, S. Wakino, S. Tatematsu, K. Yoshioka, K. Homma, N. Sugano, M. Kimoto, K. Hayashi, and H. Itoh Role of Asymmetric Dimethylarginine in Vascular Injury in Transgenic Mice Overexpressing Dimethylarginie Dimethylaminohydrolase 2 Circ. Res., July 20, 2007; 101(2): e2 - e10. [Abstract] [Full Text] [PDF]

				H. Konishi, K. Sydow, and J. P. Cooke Dimethylarginine Dimethylaminohydrolase Promotes Endothelial Repair After Vascular Injury J. Am. Coll. Cardiol., March 13, 2007; 49(10): 1099 - 1105. [Abstract] [Full Text] [PDF]

				J. Leiper and P. Vallance New tricks from an old dog: nitric oxide-independent effects of dimethylarginine dimethylaminohydrolase. Arterioscler Thromb Vasc Biol, July 1, 2006; 26(7): 1419 - 1420. [Full Text] [PDF]