Nitric oxide inhibits Oct-1 DNA binding activity in cultured vascular smooth muscle cells

Xiao Ke Liu, Darrell R. Abernethy, Nabil S. Andrawis

Research output: Contribution to journalArticle

Abstract

Since Oct-1 is a ubiquitous DNA binding protein shown to play an important role in regulating cell proliferation and possess structural characteristics consistent with a nitric oxide (NO) target, we studied NO regulation of the DNA binding activity of Oct-1 in the A7R5 vascular smooth muscle cell (VSMC) line. Two NO donors, sodium nitroprusside (SNP) and S-nitrosoacetyl-penicillamine (SNAP) were directly added to the nuclear extract-oligonucleotide reaction mixture, respectively and the effect on Oct-1 DNA binding activity was evaluated by gel shift assay. Both NO donors (0.01-1 mM) inhibited the DNA binding activity of Oct-1. This inhibitory effect was not attenuated by dithiothreitol (DTT) (1 mM) while in contrast, DTT did antagonize the effect of diamide on Oct-1 DNA binding activity. The NO effect on Oct-1 has some specificity; as the NO donors had no effect on myc DNA binding activity. The inhibitory effect of NO donors was reproduced in A7R5 cells, without affecting their viability. These findings provide the first evidence that NO inhibits the DNA binding activity of Oct-1, probably through a cGMP independent mechanism and suggests that NO may inhibit mitogenesis in part through an effect on Oct-1 DNA binding activity in VSMCs.

Original languageEnglish (US)
Pages (from-to)739-749
Number of pages11
JournalLife Sciences
Volume62
Issue number8
DOIs
StatePublished - Jan 16 1998
Externally publishedYes

Fingerprint

Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Nitric Oxide
Cells
Nitric Oxide Donors
DNA
Dithiothreitol
Diamide
Penicillamine
DNA-Binding Proteins
Cell proliferation
Nitroprusside
Oligonucleotides
Assays
Gels
Cell Proliferation
Cell Line

Keywords

  • DNA binding activity
  • Nitric oxide
  • Oct-1
  • Vascular smooth muscle cell

ASJC Scopus subject areas

  • Pharmacology

Cite this

Nitric oxide inhibits Oct-1 DNA binding activity in cultured vascular smooth muscle cells. / Liu, Xiao Ke; Abernethy, Darrell R.; Andrawis, Nabil S.

In: Life Sciences, Vol. 62, No. 8, 16.01.1998, p. 739-749.

Research output: Contribution to journalArticle

Liu, Xiao Ke ; Abernethy, Darrell R. ; Andrawis, Nabil S. / Nitric oxide inhibits Oct-1 DNA binding activity in cultured vascular smooth muscle cells. In: Life Sciences. 1998 ; Vol. 62, No. 8. pp. 739-749.
@article{e0e3bdf9e00c4717a23bae9d523d216e,
title = "Nitric oxide inhibits Oct-1 DNA binding activity in cultured vascular smooth muscle cells",
abstract = "Since Oct-1 is a ubiquitous DNA binding protein shown to play an important role in regulating cell proliferation and possess structural characteristics consistent with a nitric oxide (NO) target, we studied NO regulation of the DNA binding activity of Oct-1 in the A7R5 vascular smooth muscle cell (VSMC) line. Two NO donors, sodium nitroprusside (SNP) and S-nitrosoacetyl-penicillamine (SNAP) were directly added to the nuclear extract-oligonucleotide reaction mixture, respectively and the effect on Oct-1 DNA binding activity was evaluated by gel shift assay. Both NO donors (0.01-1 mM) inhibited the DNA binding activity of Oct-1. This inhibitory effect was not attenuated by dithiothreitol (DTT) (1 mM) while in contrast, DTT did antagonize the effect of diamide on Oct-1 DNA binding activity. The NO effect on Oct-1 has some specificity; as the NO donors had no effect on myc DNA binding activity. The inhibitory effect of NO donors was reproduced in A7R5 cells, without affecting their viability. These findings provide the first evidence that NO inhibits the DNA binding activity of Oct-1, probably through a cGMP independent mechanism and suggests that NO may inhibit mitogenesis in part through an effect on Oct-1 DNA binding activity in VSMCs.",
keywords = "DNA binding activity, Nitric oxide, Oct-1, Vascular smooth muscle cell",
author = "Liu, {Xiao Ke} and Abernethy, {Darrell R.} and Andrawis, {Nabil S.}",
year = "1998",
month = "1",
day = "16",
doi = "10.1016/S0024-3205(97)01172-7",
language = "English (US)",
volume = "62",
pages = "739--749",
journal = "Life Sciences",
issn = "0024-3205",
publisher = "Elsevier Inc.",
number = "8",

}

TY - JOUR

T1 - Nitric oxide inhibits Oct-1 DNA binding activity in cultured vascular smooth muscle cells

AU - Liu, Xiao Ke

AU - Abernethy, Darrell R.

AU - Andrawis, Nabil S.

PY - 1998/1/16

Y1 - 1998/1/16

N2 - Since Oct-1 is a ubiquitous DNA binding protein shown to play an important role in regulating cell proliferation and possess structural characteristics consistent with a nitric oxide (NO) target, we studied NO regulation of the DNA binding activity of Oct-1 in the A7R5 vascular smooth muscle cell (VSMC) line. Two NO donors, sodium nitroprusside (SNP) and S-nitrosoacetyl-penicillamine (SNAP) were directly added to the nuclear extract-oligonucleotide reaction mixture, respectively and the effect on Oct-1 DNA binding activity was evaluated by gel shift assay. Both NO donors (0.01-1 mM) inhibited the DNA binding activity of Oct-1. This inhibitory effect was not attenuated by dithiothreitol (DTT) (1 mM) while in contrast, DTT did antagonize the effect of diamide on Oct-1 DNA binding activity. The NO effect on Oct-1 has some specificity; as the NO donors had no effect on myc DNA binding activity. The inhibitory effect of NO donors was reproduced in A7R5 cells, without affecting their viability. These findings provide the first evidence that NO inhibits the DNA binding activity of Oct-1, probably through a cGMP independent mechanism and suggests that NO may inhibit mitogenesis in part through an effect on Oct-1 DNA binding activity in VSMCs.

AB - Since Oct-1 is a ubiquitous DNA binding protein shown to play an important role in regulating cell proliferation and possess structural characteristics consistent with a nitric oxide (NO) target, we studied NO regulation of the DNA binding activity of Oct-1 in the A7R5 vascular smooth muscle cell (VSMC) line. Two NO donors, sodium nitroprusside (SNP) and S-nitrosoacetyl-penicillamine (SNAP) were directly added to the nuclear extract-oligonucleotide reaction mixture, respectively and the effect on Oct-1 DNA binding activity was evaluated by gel shift assay. Both NO donors (0.01-1 mM) inhibited the DNA binding activity of Oct-1. This inhibitory effect was not attenuated by dithiothreitol (DTT) (1 mM) while in contrast, DTT did antagonize the effect of diamide on Oct-1 DNA binding activity. The NO effect on Oct-1 has some specificity; as the NO donors had no effect on myc DNA binding activity. The inhibitory effect of NO donors was reproduced in A7R5 cells, without affecting their viability. These findings provide the first evidence that NO inhibits the DNA binding activity of Oct-1, probably through a cGMP independent mechanism and suggests that NO may inhibit mitogenesis in part through an effect on Oct-1 DNA binding activity in VSMCs.

KW - DNA binding activity

KW - Nitric oxide

KW - Oct-1

KW - Vascular smooth muscle cell

UR - http://www.scopus.com/inward/record.url?scp=0032535947&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032535947&partnerID=8YFLogxK

U2 - 10.1016/S0024-3205(97)01172-7

DO - 10.1016/S0024-3205(97)01172-7

M3 - Article

C2 - 9489510

AN - SCOPUS:0032535947

VL - 62

SP - 739

EP - 749

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

IS - 8

ER -