The superoxide‐producing NAD(P)H oxidase Nox4 in the nucleus of human vascular endothelial cells

Genes to Cells - Tập 10 Số 12 - Trang 1139-1151 - 2005
Junya Kuroda1, Kazunori Nakagawa2, Tomoko Yamasaki1, Kei‐ichiro Nakamura3, Ryu Takeya4,1, Futoshi Kuribayashi1, Shinobu Imajoh‐Ohmi5, Kazuhiko Igarashi4,6, Yosaburo Shibata3, Katsuo Sueishi2, Hideki Sumimoto4,1
1Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan
2Department of Pathology and
3Department of Developmental Molecular Anatomy, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan
4CREST, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
5Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan
6Department of Biomedical Chemistry, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima 734-8551, Japan

Tóm tắt

The superoxide‐producing NAD(P)H oxidase Nox4 was initially identified as an enzyme that is highly expressed in the kidney and is possibly involved in oxygen sensing and cellular senescence. Although the oxidase is also abundant in vascular endothelial cells, its role remains to be elucidated. Here we show that Nox4 preferentially localizes to the nucleus of human umbilical vein endothelial cells (HUVECs), by immunocytochemistry and immunoelectron microscopy using three kinds of affinity‐purified antibodies raised against distinct immunogens from human Nox4. Silencing of Nox4 by RNA interference (RNAi) abrogates nuclear signals given with the antibodies, confirming the nuclear localization of Nox4. The nuclear fraction of HUVECs exhibits an NAD(P)H‐dependent superoxide‐producing activity in a manner dependent on Nox4, which activity can be enhanced upon cell stimulation with phorbol 12‐myristate 13‐acetate. This stimulant also facilitates gene expression as estimated in the present transfection assay of HUVECs using a reporter regulated by the Maf‐recognition element MARE, a DNA sequence that constitutes a part of oxidative stress response. Both basal and stimulated transcriptional activities are impaired by RNAi‐mediated Nox4 silencing. Thus Nox4 appears to produce superoxide in the nucleus of HUVECs, thereby regulating gene expression via a mechanism for oxidative stress response.

Từ khóa


Tài liệu tham khảo

10.1161/01.CIR.0000105680.92873.70

10.1074/jbc.274.25.18055

10.1093/nar/gkg251

10.1074/jbc.M406486200

10.1074/jbc.C200613200

10.1126/science.287.5450.138

10.1074/jbc.M103034200

10.1113/jphysiol.2003.055913

10.1016/S0968-0004(03)00194-4

10.1038/sj.onc.1206654

10.1161/01.ATV.19.1.131

10.1016/S0378-1119(01)00449-8

10.1074/jbc.M400660200

10.1016/S0005-2728(98)00029-2

10.1152/physrev.00018.2001

10.1038/35078107

10.1007/s00125-003-1205-6

10.1073/pnas.130135897

10.1074/jbc.M301289200

10.1074/jbc.R400024200

10.1152/ajprenal.00414.2002

10.1136/hrt.2003.029397

10.1016/S0076-6879(96)68040-9

10.1016/j.freeradbiomed.2004.08.011

10.1161/01.ATV.0000112024.13727.2c

10.1074/jbc.275.20.15370

10.1038/367568a0

10.1016/S0021-9258(18)48476-1

10.1242/jcs.114.23.4253

10.1101/gad.13.1.76

10.1074/jbc.274.35.25051

10.1093/emboj/cdf642

10.1038/nri1312

10.1152/ajpregu.00758.2002

10.1074/jbc.M110073200

10.1007/s00418-004-0679-8

10.1101/gad.1172504

10.1006/expr.1993.1102

10.1126/stke.2000.53.pe1

10.1073/pnas.89.19.9176

10.1074/jbc.M007597200

10.1038/43459

10.1016/j.bbrc.2005.08.210

10.1161/hq0102.102189

10.1074/jbc.M212856200

10.1074/jbc.M414548200

10.1126/science.1069300

Thông tin
Thông tin xuất bản

Genes to Cells

Tập 10 Số 12

1139-1151

Thông tin tác giả