TY - JOUR
T1 - The human preproendothelin-1 gene
T2 - Possible regulation by endothelial phosphoinositide turnover signaling
AU - Yanagisawa, Masashi
AU - Inoue, A.
AU - Takuwa, Y.
AU - Mitsui, Y.
AU - Kobayashi, M.
AU - Masaki, T.
PY - 1989
Y1 - 1989
N2 - Preproendothelin-1 (ppET-1) mRNA has previously been demonstrated to be markedly induced in cultured endothelial cells (EC) by the addition to the medium of thrombin, an agent known to stimulate phosphoinositide turnover in EC. In this study, the mechanism of regulation of ppET-1 mRNA expression was investigated in cultured human umbilical vein EC by RNA blot analysis with cloned ppET-1 gene as a probe. The mRNA for ppET-1 was rapidly upregulated by O-tetradecanoylphorbol-13-acetate (TPA) (0.5 µM) and by ionomycin (5 µM) within 10 min of addition to the medium, but not by forskolin (50 µM). The rapid induction of ppET-1 mRNA by TPA or ionomycin occurred even in the presence of cycloheximide, indicating that the mRNA induction does not require de novo protein synthesis. The ppET-1 mRNA was an extremely unstable species of mRNA with an apparent half-life of about 15 min. However, the half-life of ppET-1 mRNA was not appreciably affected by TPA or ionomycin, suggesting that the mRNA induction by these agents is mostly due to an activation of the transcription of the mRNA. These observations indicate that the production of ET-1 in human EC can be controlled by a transcriptional gene regulation directly coupled to the intracellular signals from the phosphoinositide-turnover pathway, i.e., activation of protein kinase C and increase in intracellular Ca2+. These mechanisms are discussed in relation to information on the primary structure of cloned ppET-1 gene.
AB - Preproendothelin-1 (ppET-1) mRNA has previously been demonstrated to be markedly induced in cultured endothelial cells (EC) by the addition to the medium of thrombin, an agent known to stimulate phosphoinositide turnover in EC. In this study, the mechanism of regulation of ppET-1 mRNA expression was investigated in cultured human umbilical vein EC by RNA blot analysis with cloned ppET-1 gene as a probe. The mRNA for ppET-1 was rapidly upregulated by O-tetradecanoylphorbol-13-acetate (TPA) (0.5 µM) and by ionomycin (5 µM) within 10 min of addition to the medium, but not by forskolin (50 µM). The rapid induction of ppET-1 mRNA by TPA or ionomycin occurred even in the presence of cycloheximide, indicating that the mRNA induction does not require de novo protein synthesis. The ppET-1 mRNA was an extremely unstable species of mRNA with an apparent half-life of about 15 min. However, the half-life of ppET-1 mRNA was not appreciably affected by TPA or ionomycin, suggesting that the mRNA induction by these agents is mostly due to an activation of the transcription of the mRNA. These observations indicate that the production of ET-1 in human EC can be controlled by a transcriptional gene regulation directly coupled to the intracellular signals from the phosphoinositide-turnover pathway, i.e., activation of protein kinase C and increase in intracellular Ca2+. These mechanisms are discussed in relation to information on the primary structure of cloned ppET-1 gene.
KW - Ca ionophore
KW - Endothelial cells
KW - Human
KW - Phorbol ester
KW - Phosphoinositide turnover
KW - Preproendothelin-1 gene
UR - http://www.scopus.com/inward/record.url?scp=0024583007&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0024583007&partnerID=8YFLogxK
U2 - 10.1097/00005344-198900135-00005
DO - 10.1097/00005344-198900135-00005
M3 - Article
C2 - 2473287
AN - SCOPUS:0024583007
SN - 0160-2446
VL - 13
SP - S13-S17
JO - Journal of Cardiovascular Pharmacology
JF - Journal of Cardiovascular Pharmacology
ER -