Abstract
Two major troponin I (TnI) genes, fetal TnI (ssTnI) and adult TnI (cTnI), are expressed in the mammalian heart under the control of a developmentally regulated program. In this study, the up-stream domain (∼1,800 bp) of mouse fetal TnI gene has been cloned and characterized. There is a high homology of this region among mouse, rat and human. Analysis of the sequence revealed several putative regulatory domains and binding sites (Sp1 binding sites, GATA binding site, MyoD, CREB, MEF2, AP1, NFκB, etc). Transfection assays indicated that conserved GA-rich sequences, CREB and a CCAAT box within the first 300 bp upstream of the transcription start site were critical for the gene expression. Electrophoretic mobility shift assays (EMSAs) and chromatin immunoprecipitation (ChIP) assays revealed binding proteins to CREB site in nuclear extracts from myocardial cells. An inhibitory domain was revealed within the sequence between -1,700 to -1,780. Thyroid hormone (T3) caused a significant inhibitory effect on ssTnI expression in myocardial cells whereas this effect was not evident in CHO cells.
| Original language | English |
|---|---|
| Pages (from-to) | 605-613 |
| Number of pages | 9 |
| Journal | Journal of Biomedical Science |
| Volume | 15 |
| Issue number | 5 |
| DOIs | |
| State | Published - Sep 1 2008 |
| Externally published | Yes |
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Keywords
- Gene regulation
- Myocardium
- Myofibril protein
- Slow skeletal troponin I
- Thyroid hormone
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Functional characterization of mouse fetal TnI gene promoters in myocardial cells. / Du, J.; Nan, C.; Huang, J. J.; Zhang, Chi; Liu, J.; Jia, P.; Abers, M.; Huang, X. P.
In: Journal of Biomedical Science, Vol. 15, No. 5, 01.09.2008, p. 605-613.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Functional characterization of mouse fetal TnI gene promoters in myocardial cells
AU - Du, J.
AU - Nan, C.
AU - Huang, J. J.
AU - Zhang, Chi
AU - Liu, J.
AU - Jia, P.
AU - Abers, M.
AU - Huang, X. P.
PY - 2008/9/1
Y1 - 2008/9/1
N2 - Two major troponin I (TnI) genes, fetal TnI (ssTnI) and adult TnI (cTnI), are expressed in the mammalian heart under the control of a developmentally regulated program. In this study, the up-stream domain (∼1,800 bp) of mouse fetal TnI gene has been cloned and characterized. There is a high homology of this region among mouse, rat and human. Analysis of the sequence revealed several putative regulatory domains and binding sites (Sp1 binding sites, GATA binding site, MyoD, CREB, MEF2, AP1, NFκB, etc). Transfection assays indicated that conserved GA-rich sequences, CREB and a CCAAT box within the first 300 bp upstream of the transcription start site were critical for the gene expression. Electrophoretic mobility shift assays (EMSAs) and chromatin immunoprecipitation (ChIP) assays revealed binding proteins to CREB site in nuclear extracts from myocardial cells. An inhibitory domain was revealed within the sequence between -1,700 to -1,780. Thyroid hormone (T3) caused a significant inhibitory effect on ssTnI expression in myocardial cells whereas this effect was not evident in CHO cells.
AB - Two major troponin I (TnI) genes, fetal TnI (ssTnI) and adult TnI (cTnI), are expressed in the mammalian heart under the control of a developmentally regulated program. In this study, the up-stream domain (∼1,800 bp) of mouse fetal TnI gene has been cloned and characterized. There is a high homology of this region among mouse, rat and human. Analysis of the sequence revealed several putative regulatory domains and binding sites (Sp1 binding sites, GATA binding site, MyoD, CREB, MEF2, AP1, NFκB, etc). Transfection assays indicated that conserved GA-rich sequences, CREB and a CCAAT box within the first 300 bp upstream of the transcription start site were critical for the gene expression. Electrophoretic mobility shift assays (EMSAs) and chromatin immunoprecipitation (ChIP) assays revealed binding proteins to CREB site in nuclear extracts from myocardial cells. An inhibitory domain was revealed within the sequence between -1,700 to -1,780. Thyroid hormone (T3) caused a significant inhibitory effect on ssTnI expression in myocardial cells whereas this effect was not evident in CHO cells.
KW - Gene regulation
KW - Myocardium
KW - Myofibril protein
KW - Slow skeletal troponin I
KW - Thyroid hormone
UR - http://www.scopus.com/inward/record.url?scp=51349117727&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=51349117727&partnerID=8YFLogxK
U2 - 10.1007/s11373-008-9246-y
DO - 10.1007/s11373-008-9246-y
M3 - Article
VL - 15
SP - 605
EP - 613
JO - Journal of Biomedical Science
JF - Journal of Biomedical Science
SN - 1021-7770
IS - 5
ER -