Prostaglandin F2alpha inhibits SERCA2 gene transcription through an induction of Egr-1 in cultured neonatal rat cardiac myocytes.
Prostaglandin F(2alpha) (PGF(2alpha)) stimulates hypertrophic growth of neonatal rat cardiac myocytes, a feature of which includes downregulation of the Ca(2+)-ATPase (SERCA2), a major Ca(2+) transport protein in SR. The molecular mechanisms by which PGF(2alpha) inhibits SERCA2 gene expression remain unknown. We determined the cis-regulatory elements responsible for the regulation of the SERCA2 gene expression in cultured neonatal rat cardiac myocytes exposed to PGF(2alpha). The role of Egr-1 was evaluated by transient transfection of its expression vector and antisense oligonucleotide. Signaling pathways were determined by using the pharmacological inhibitors or cDNA expression plasmids coding for dominant negative forms of Ras and Rac. PGF(2alpha) reduced the SERCA2 mRNA levels in a time- and dose-dependent manner in cultured rat cardiac myocytes. Transient transfection analyses showed that PGF(2alpha) -responsive elements are located between -284 and -72 of the SERCA2 promoter, which contains G+C-rich sequences homologous to Sp1, Egr-1 and AP2-binding sites. PGF(2alpha) significantly increased Egr-1 expression, and overexpression of Egr-1 largely reduced the transcription of the SERCA2 gene. Egr-1 antisense oligonucleotides blocked the PGF(2alpha) -mediated decrease in SERCA2 mRNA expression. Furthermore, inhibitors for either genistein-sensitive tyrosine kinase or p38 MAPK, and dominant negative forms of either Ras or Rac, prevented PGF(2alpha) -induced repression of SERCA2 mRNA levels. These results suggest that Egr-1, as well as Ras, Rac, and p38 MAPK, plays a crucial role in the repression of SERCA2 gene expression during PGF(2alpha) -induced cardiac hypertrophy.
Hara
Shiro
S
Department of Medicine and Biological Science, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
Arai
Masashi
M
Tomaru
Koichi
K
Doi
Hiroshi
H
Koitabashi
Norimichi
N
Iso
Tatsuya
T
Watanabe
Atai
A
Tanaka
Toru
T
Maeno
Toshitaka
T
Suga
Tatsuo
T
Yokoyama
Tomoyuki
T
Kurabayashi
Masahiko
M
D000818:Animals; D002478:Cells, Cultured; D015237:Dinoprost / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects*; D051766:Early Growth Response Protein 1 / Q000096:biosynthesis*; D015870:Gene Expression; D032383:Myocytes, Cardiac / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism*; D014158:Transcription, Genetic
C506948:Atp2a2 protein, rat; D051766:Early Growth Response Protein 1; C486110:Egr1 protein, rat; D015237:Dinoprost; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases