A novel TCTG(G/C) direct repeat and an A/T-rich HMG2-binding site control the expression of the rat cardiac troponin T gene

Qinchuan Wang, Jenny Li Chun Lin, Jim Jung Ching Lin

Research output: Contribution to journalArticle

Abstract

We previously showed that the rat cardiac troponin T proximal promoter (-497 bp from the transcriptional start site) was sufficient to confer cardiac-specific expression of a reporter gene in both cultured cardiomyocytes and transgenic mice. This promoter consists of two cis-regulatory modules with high sequence similarity. Nucleotides from -319 to -289 (termed D2) and nucleotides from -249 to -209 (termed F41) contain a TCTG(G/C) direct repeat and an A/T-rich sequence. Competition gel mobility shift assays revealed that the same protein factors were bound to both D2 and F41. Deletion and religation studies of the promoter suggested that D2 acted as an enhancer but could not totally substitute for the F41 function. Mutational analyses demonstrated that the direct repeat was required for the DNA-binding and promoter activity. Moreover, cardiac-specific 42 kDa proteins and a ubiquitous high mobility group 2 (HMG2) protein were identified to be responsible for the binding to the TCTG(G/C) direct repeat and the A/T-rich sequence, respectively. Overexpression of HMG2 had differential effects on the promoter in cardiomyocytes versus fibroblasts. Our results provided the first evidence to support that HMG2 together with tissue-specific factors could direct a stimulatory or inhibitory effect on the cardiac troponin T gene expression in heart or non-heart tissue, respectively.

Original languageEnglish (US)
Pages (from-to)1667-1679
Number of pages13
JournalJournal of Molecular and Cellular Cardiology
Volume34
Issue number12
DOIs
StatePublished - Dec 1 2002
Externally publishedYes

Fingerprint

Troponin T
Nucleic Acid Repetitive Sequences
Binding Sites
Cardiac Myocytes
Nucleotides
High Mobility Group Proteins
Genes
Thromboplastin
Electrophoretic Mobility Shift Assay
Reporter Genes
Transgenic Mice
Proteins
Fibroblasts
Gels
Gene Expression
DNA

Keywords

  • A/T-rich sequence
  • Gel mobility shift assay
  • HMG2
  • Promoter
  • TCTG(G/C) direct repeat

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

A novel TCTG(G/C) direct repeat and an A/T-rich HMG2-binding site control the expression of the rat cardiac troponin T gene. / Wang, Qinchuan; Lin, Jenny Li Chun; Lin, Jim Jung Ching.

In: Journal of Molecular and Cellular Cardiology, Vol. 34, No. 12, 01.12.2002, p. 1667-1679.

Research output: Contribution to journalArticle

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