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 Honma, Naoki Sugano, Masahiko Kurabayashi, Takao Saruta, Koichi Hayashi

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1488-1494
Number of pages7
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume26
Issue number7
DOIs
Publication statusPublished - 2006 Jul

Fingerprint

Sp1 Transcription Factor
Vascular Endothelial Growth Factor A
Endothelial Cells
Cyclic AMP-Dependent Protein Kinases
Nitric Oxide Synthase
Transfection
Human Umbilical Vein Endothelial Cells
Threonine
Nuclear Proteins
dimethylargininase
Oligonucleotides
Small Interfering RNA
Atherosclerosis
Protein Isoforms
Proteins
Up-Regulation
Phosphorylation
DNA
Enzymes

Keywords

  • ADMA
  • DDAH
  • Endothelial cell
  • Sp1
  • VEGF

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Dimethylarginine dimethylaminohydrolase 2 increases vascular endothelial growth factor expression through Sp1 transcription factor in endothelial cells. / Hasegawa, Kazuhiro; Wakino, Shu; Tanaka, Toru; Kimoto, Masumi; Tatematsu, Satoru; Kanda, Takeshi; Yoshioka, Kyoko; Honma, Koichiro; Sugano, Naoki; Kurabayashi, Masahiko; Saruta, Takao; Hayashi, Koichi.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 26, No. 7, 07.2006, p. 1488-1494.

Research output: Contribution to journalArticle

Hasegawa, Kazuhiro ; Wakino, Shu ; Tanaka, Toru ; Kimoto, Masumi ; Tatematsu, Satoru ; Kanda, Takeshi ; Yoshioka, Kyoko ; Honma, Koichiro ; Sugano, Naoki ; Kurabayashi, Masahiko ; Saruta, Takao ; Hayashi, Koichi. / Dimethylarginine dimethylaminohydrolase 2 increases vascular endothelial growth factor expression through Sp1 transcription factor in endothelial cells. In: Arteriosclerosis, Thrombosis, and Vascular Biology. 2006 ; Vol. 26, No. 7. pp. 1488-1494.
@article{02077f6e6a4f4dbb8ab10c26eaa0010e,
title = "Dimethylarginine dimethylaminohydrolase 2 increases vascular endothelial growth factor expression through Sp1 transcription factor in endothelial cells",
abstract = "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.",
keywords = "ADMA, DDAH, Endothelial cell, Sp1, VEGF",
author = "Kazuhiro Hasegawa and Shu Wakino and Toru Tanaka and Masumi Kimoto and Satoru Tatematsu and Takeshi Kanda and Kyoko Yoshioka and Koichiro Honma and Naoki Sugano and Masahiko Kurabayashi and Takao Saruta and Koichi Hayashi",
year = "2006",
month = "7",
doi = "10.1161/01.ATV.0000219615.88323.b4",
language = "English",
volume = "26",
pages = "1488--1494",
journal = "Arteriosclerosis, Thrombosis, and Vascular Biology",
issn = "1079-5642",
publisher = "Lippincott Williams and Wilkins",
number = "7",

}

TY - JOUR

T1 - Dimethylarginine dimethylaminohydrolase 2 increases vascular endothelial growth factor expression through Sp1 transcription factor in endothelial cells

AU - Hasegawa, Kazuhiro

AU - Wakino, Shu

AU - Tanaka, Toru

AU - Kimoto, Masumi

AU - Tatematsu, Satoru

AU - Kanda, Takeshi

AU - Yoshioka, Kyoko

AU - Honma, Koichiro

AU - Sugano, Naoki

AU - Kurabayashi, Masahiko

AU - Saruta, Takao

AU - Hayashi, Koichi

PY - 2006/7

Y1 - 2006/7

N2 - 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.

AB - 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.

KW - ADMA

KW - DDAH

KW - Endothelial cell

KW - Sp1

KW - VEGF

UR - http://www.scopus.com/inward/record.url?scp=33745953244&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745953244&partnerID=8YFLogxK

U2 - 10.1161/01.ATV.0000219615.88323.b4

DO - 10.1161/01.ATV.0000219615.88323.b4

M3 - Article

C2 - 16574895

AN - SCOPUS:33745953244

VL - 26

SP - 1488

EP - 1494

JO - Arteriosclerosis, Thrombosis, and Vascular Biology

JF - Arteriosclerosis, Thrombosis, and Vascular Biology

SN - 1079-5642

IS - 7

ER -