Transcription of the SERCA2 Gene is Decreased in Pressure-overloaded Hearts: A Study Using In Vivo Direct Gene Transfer into Living Myocardium

doi:10.1006/jmcc.1999.1045

Journal of Molecular and Cellular Cardiology

Volume 31, Issue 12, December 1999, Pages 2167-2174

https://doi.org/10.1006/jmcc.1999.1045 Get rights and content

Abstract

T. Takizawa, M. Arai, A. Yoguchi, K. Tomaru, M. Kurabayashi and R. Nagai. Transcription of the SERCA2 Gene is Decreased in Pressure-overloaded Hearts: A Study Using In Vivo Direct Gene Transfer into Living Myocardium.Journal of Molecular and Cellular Cardiology (1999) 31, 2167–2174. The sarcoplasmic reticulum Ca²⁺-ATPase (SERCA2) controls the myocardial relaxation process. Under pressure-overload, the expression of its mRNA decreases, thus controlling cardiac function to conform to the load. However, it is not known whether this decreased expression is caused by a decrease in the transcription of the SERCA2 gene. The object of this study was to determine the transcription control mechanism of the SERCA2 gene under pressure-overload in vivo, and to identify the pressure-overload-sensitive regions of the SERCA2 gene. Ten micrograms of a plasmid, containing the 5′ upstream (−1810 bp to +350 bp) region of the SERCA2 gene and a luciferase reporter gene, were introduced into adult rat myocardium by in vivo direct gene transfer, and the luciferase activity was measured 5 days later. The transcriptional activity under pressure-overload decreased to 27±17% of the control. Based on this result, we concluded that the decreased mRNA expression of SERCA2 in pressure-overload cardiac hypertrophy is due to decreased gene transcription. In addition, various deletion fragments of the SERCA2 promoter region were produced, and tested for luciferase production under pressure-overload. Our data suggest that a transcription activation site is present between −685 and −284 bp, and two transcription inhibition sites are present between −1810 to −1110 bp and −284 to −72 bp. These may be the pressure-sensitive regions of the SERCA2 gene of in vivo hypertrophied myocardium under pressure-overload.

References (22)

M Arai et al.
Sarcoplasmic reticulum genes are upregulated in mild cardiac hypertrophy but downregulated in severe cardiac hypertrophy induced by pressure-overload
J Mol Cell Cardiol
(1996)
DL Baker et al.
Multiple SP1 binding sites in the cardiac/slow twitch muscle sarcoplasmic reticulum Ca²⁺-ATPase gene promoter are required for expression in Sol8 muscle cells
J Biol Chem
(1996)
P Chomczynski et al.
Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction
Anal Biochem
(1987)
M Arai et al.
Sarcoplasmic reticulum gene expression in cardiac hypertrophy and heart failure
Circ Res
(1994)
D de la Bastie et al.
Function of the sarcoplasmic reticulum and expression of its Ca²⁺-ATPase gene in pressure-overload-induced cardiac hypertrophy in the rat
Circ Res
(1990)
PM Buttrick et al.
Behavior of genes directly injected into the rat heart in vivo
Circ Res
(1992)
T Cornelius et al.
Regulation of the rat atrial natriuretic peptide gene after acute imposition of left ventricular pressure-overload
Hypertension
(1997)
CR Cory et al.
Role of sarcoplasmic reticulum in loss of load-sensitive relaxation in pressure-overload cardiac hypertrophy
Am J Physiol
(1994)
AM Feldman et al.
Selective changes in cardiac gene expression during compensated hypertrophy and the transition to cardiac decompensation in rats with chronic aortic banding
Circ Res
(1993)
SA Fisher et al.
Characterization of promoter elements of the rabbit cardiac sarcoplasmic reticulum Ca²⁺-ATPase gene required for expression in cardiac muscle cells
Circ Res
(1993)

R Harsdorf et al.

Identification of a cis-acting regulatory element conferring inducibility of the atrial natriuretic factor gene in acute pressure-overload

J Clin Invest

(1997)

Cited by (26)

Mechanical stress-dependent transcriptional regulation of sarcolipin gene in the rodent atrium
2005, Biochemical and Biophysical Research Communications
Citation Excerpt :
We found that the expression level of SERCA2a mRNA was also decreased in the atria by mechanical stress. Two previous studies have demonstrated that the transcription of SERCA2a in the atria is decreased by mechanical stress [4,5], which is consistent with data form many studies investigating the change in SERCA2a transcripts in the pressure or volume-overloaded ventricles [2,18–20]. Although we did not assess the functional consequences of reduction in the SERCA2a transcripts by mechanical stress, a considerable number of studies have demonstrated that the reduction in SERCA2a mRNA results in a subsequent decrease in SERCA2a proteins and Ca2+ uptake in ventricles with mechanical stress [21–23].
Sarcolipin, a homologue of phospholamban, regulates Ca²⁺ uptake through the interaction with sarcoplasmic reticulum Ca²⁺ATPase (SERCA) and is predominantly expressed in the atrial muscle. Although the atrial chamber-specific expression of sarcolipin could be primarily regulated at the transcriptional level, the transcriptional regulation remains poorly understood. Since mechanical stress plays an important role in transcriptional regulation of a gene involved in cardiac hypertrophy and remodeling, we generated left-sided or right-sided pressure-overload models by transverse aortic constriction (TAC) in ddY mice or by monocrotaline administration in Wistar rats, respectively. TAC significantly decreased the expression of sarcolipin, SERCA2a, and phospholamban mRNAs in the left atrium (LA) than those in the right atrium (RA). By contrast, monocrotaline administration significantly decreased the expression of sarcolipin, SERCA2a, and phospholamban mRNAs in the RA than those in the LA. The two independent complementary experiments unequivocally demonstrated that mechanical stress down-regulates the transcription of the sarcolipin gene.
Phenotypes of SERCA and PMCA knockout mice
2004, Biochemical and Biophysical Research Communications
Citation Excerpt :
The major objectives were to determine whether loss of one copy of the SERCA2 gene would lead to impaired cardiac performance or heart disease and whether compensatory mechanisms were activated in the heart. This is of interest because many studies have shown that SERCA2a mRNA, protein, and activity are reduced in diseased cardiac muscle and it has been suggested that these reductions may contribute to the pathogenic process [38–40]. Given the role of SERCA2a as the cardiac SR Ca2+ pump and SERCA2b as the major ER Ca2+ pump in all tissues, it seemed unlikely that null mutants would survive to birth.
P-type Ca²⁺-ATPases of the sarco(endo)plasmic reticulum (SERCAs) and plasma membrane (PMCAs) are responsible for maintaining the Ca²⁺ gradients across cellular membranes that are required for regulation of Ca²⁺-mediated signaling and other biological processes. Gene-targeting studies of SERCA isoforms 1, 2, and 3 and PMCA isoforms 1, 2, and 4 have confirmed some of the general functions proposed for these pumps, such as a major role in excitation–contraction coupling for SERCA1 and SERCA2 and housekeeping functions for PMCA1 and SERCA2, but have also revealed some unexpected phenotypes. These include squamous cell cancer and plasticity in the regulation of Ca²⁺-mediated exocytosis in SERCA2 heterozygous mutant mice, modulation of Ca²⁺ signaling in SERCA3-deficient mice, deafness and balance disorders in PMCA2 null mice, and male infertility in PMCA4 null mice. These unique phenotypes provide new information about the cellular functions of these pumps, the requirement of their activities for higher order physiological processes, and the pathophysiological consequences of pump dysfunction.
Transcription factor Sp1 regulates SERCA2 gene expression in pressure-overloaded hearts: A study using in vivo direct gene transfer into living myocardium
2003, Journal of Molecular and Cellular Cardiology
Pressure-overload hypertrophy results in downregulation of the sarcoplasmic reticulum Ca²⁺-ATPase pump encoding SERCA2 gene that regulates Ca²⁺ uptake and myocardial relaxation. We previously characterized a proximal promoter region containing four Sp1 element consensus sequences (–284 to –72 base pairs (bp)) that was responsible for pressure-overload-induced transcriptional regulation. The purpose of the present study was to determine which of the Sp1 sites was responsible for the downregulation of SERCA2 gene transcription under pressure overload. Using an in vivo direct gene transfer assay, SERCA2 gene transcriptional activity was measured under pressure overload. Site-directed mutagenesis of the four Sp1 sites (I-IV) in the SERCA2 gene promoter (–284 to –72 bp) was performed. Wild-type and Sp1 mutant-luciferase reporter constructs were injected into the left-ventricular apices of pressure overload or sham-operated rats, and Sp1 mRNA and SERCA2 gene-luciferase activity was measured sequentially from 3 to 14 d after surgery. At 5 d, Sp1 mRNA in the pressure-overload rats increased to 124 ± 7% of sham group levels, and pressure-overload-induced SERCA2 transcriptional activity was 15 ± 4% of sham group when all four Sp1 sites remained intact. Mutation of the Sp1 mutant sites I (–196 to –191 bp) and III (–118 to –113 bp) blocked the inhibitory effect of pressure overload and resulted in SERCA2 gene transcriptional activity of 54 ± 15% and 56 ± 7% of sham group, respectively. We conclude that the pressure-overload-induced decrease in SERCA2 mRNA is mediated by Sp1 sites I and III.
Advanced glycation endproducts and their receptor: Do they play a role in diabetic cardiomyopathy?
2002, Journal of Molecular and Cellular Cardiology
Left ventricular SERCA2a gene down-regulation does not parallel ANP gene up-regulation during post-MI remodelling in rats
2005, European Journal of Heart Failure
Contractile arrest reveals calcium-dependent stimulation of SERCA2a mRNA expression in cultured ventricular cardiomyocytes
2004, Cardiovascular Research

View all citing articles on Scopus

^f1: Please address all correspondence to: Masashi Arai, Second Department of Internal Medicine, Gunma University School of Medicine, 3-39-15 Showa-machi, Maebashi, Gunma, 371-8511, Japan.

View full text

Article preview

Journal of Molecular and Cellular Cardiology

Abstract

References (22)

Sarcoplasmic reticulum genes are upregulated in mild cardiac hypertrophy but downregulated in severe cardiac hypertrophy induced by pressure-overload

J Mol Cell Cardiol

Multiple SP1 binding sites in the cardiac/slow twitch muscle sarcoplasmic reticulum Ca²⁺-ATPase gene promoter are required for expression in Sol8 muscle cells

J Biol Chem

Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction

Anal Biochem

Sarcoplasmic reticulum gene expression in cardiac hypertrophy and heart failure

Circ Res

Function of the sarcoplasmic reticulum and expression of its Ca²⁺-ATPase gene in pressure-overload-induced cardiac hypertrophy in the rat

Circ Res

Behavior of genes directly injected into the rat heart in vivo

Circ Res

Regulation of the rat atrial natriuretic peptide gene after acute imposition of left ventricular pressure-overload

Hypertension

Role of sarcoplasmic reticulum in loss of load-sensitive relaxation in pressure-overload cardiac hypertrophy

Am J Physiol

Selective changes in cardiac gene expression during compensated hypertrophy and the transition to cardiac decompensation in rats with chronic aortic banding

Circ Res

Characterization of promoter elements of the rabbit cardiac sarcoplasmic reticulum Ca²⁺-ATPase gene required for expression in cardiac muscle cells

Circ Res

Identification of a cis-acting regulatory element conferring inducibility of the atrial natriuretic factor gene in acute pressure-overload

J Clin Invest

Cited by (26)

Mechanical stress-dependent transcriptional regulation of sarcolipin gene in the rodent atrium

Phenotypes of SERCA and PMCA knockout mice

Transcription factor Sp1 regulates SERCA2 gene expression in pressure-overloaded hearts: A study using in vivo direct gene transfer into living myocardium

Advanced glycation endproducts and their receptor: Do they play a role in diabetic cardiomyopathy?

Left ventricular SERCA2a gene down-regulation does not parallel ANP gene up-regulation during post-MI remodelling in rats

Contractile arrest reveals calcium-dependent stimulation of SERCA2a mRNA expression in cultured ventricular cardiomyocytes

Regular ArticleTranscription of the SERCA2 Gene is Decreased in Pressure-overloaded Hearts: A Study Using In Vivo Direct Gene Transfer into Living Myocardium

Abstract

J Mol Cell Cardiol

J Biol Chem

Anal Biochem

Sarcoplasmic reticulum gene expression in cardiac hypertrophy and heart failure

Circ Res

Function of the sarcoplasmic reticulum and expression of its Ca2+-ATPase gene in pressure-overload-induced cardiac hypertrophy in the rat

Circ Res

Behavior of genes directly injected into the rat heart in vivo

Circ Res

Regulation of the rat atrial natriuretic peptide gene after acute imposition of left ventricular pressure-overload

Hypertension

Role of sarcoplasmic reticulum in loss of load-sensitive relaxation in pressure-overload cardiac hypertrophy

Am J Physiol

Selective changes in cardiac gene expression during compensated hypertrophy and the transition to cardiac decompensation in rats with chronic aortic banding

Circ Res

Characterization of promoter elements of the rabbit cardiac sarcoplasmic reticulum Ca2+-ATPase gene required for expression in cardiac muscle cells

Circ Res

Identification of a cis-acting regulatory element conferring inducibility of the atrial natriuretic factor gene in acute pressure-overload

J Clin Invest

Regular Article
Transcription of the SERCA2 Gene is Decreased in Pressure-overloaded Hearts: A Study Using In Vivo Direct Gene Transfer into Living Myocardium

Function of the sarcoplasmic reticulum and expression of its Ca²⁺-ATPase gene in pressure-overload-induced cardiac hypertrophy in the rat

Characterization of promoter elements of the rabbit cardiac sarcoplasmic reticulum Ca²⁺-ATPase gene required for expression in cardiac muscle cells