Absence of mutation at the 50-upstream promoter region of the TPM4 gene from cardiac mutant axolotl (Ambystoma mexicanum)

Christopher R. Denz, Chi Zhang, Pingping Jia, Jianfeng Du, Xupei Huang, Syamalima Dube, Anish Thomas, Bernard J. Poiesz, Dipak K. Dube

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Tropomyosins are a family of actin-binding proteins that show cell-specific diversity by a combination of multiple genes and alternative RNA splicing. Of the 4 different tropomyosin genes, TPM4 plays a pivotal role in myofibrillogenesis as well as cardiac contractility in amphibians. In this study, we amplified and sequenced the upstream regulatory region of the TPM4 gene from both normal and mutant axolotl hearts. To identify the cis-elements that are essential for the expression of the TPM4, we created various deletion mutants of the TPM4 promoter DNA, inserted the deleted segments into PGL3 vector, and performed promoter-reporter assay using luciferase as the reporter gene. Comparison of sequences of the promoter region of the TPM4 gene from normal and mutant axolotl revealed no mutations in the promoter sequence of the mutant TPM4 gene. CArG box elements that are generally involved in controlling the expression of several other muscle-specific gene promoters were not found in the upstream regulatory region of the TPM4 gene. In deletion experiments, loss of activity of the reporter gene was noted upon deletion which was then restored upon further deletion suggesting the presence of both positive and negative cis-elements in the upstream regulatory region of the TPM4 gene. We believe that this is the first axolotl promoter that has ever been cloned and studied with clear evidence that it functions in mammalian cell lines. Although striated muscle-specific cis-acting elements are absent from the promoter region of TPM4 gene, our results suggest the presence of positive and negative cis-elements in the promoter region, which in conjunction with positive and negative trans-elements may be involved in regulating the expression of TPM4 gene in a tissue-specific manner.

Original languageEnglish
Pages (from-to)235-243
Number of pages9
JournalCardiovascular Toxicology
Volume11
Issue number3
DOIs
StatePublished - Sep 1 2011
Externally publishedYes

Fingerprint

Ambystoma mexicanum
Genetic Promoter Regions
Genes
Mutation
Nucleic Acid Regulatory Sequences
Tropomyosin
Reporter Genes
Muscle
Microfilament Proteins
Striated Muscle
Muscle Development
Alternative Splicing
Amphibians
Luciferases
Gene Expression
Cell Line

Keywords

  • Ambystoma mexicanum
  • Myofibrillogenesis
  • Promoter
  • Tropomyosin

ASJC Scopus subject areas

  • Toxicology
  • Cardiology and Cardiovascular Medicine
  • Molecular Biology

Cite this

Absence of mutation at the 50-upstream promoter region of the TPM4 gene from cardiac mutant axolotl (Ambystoma mexicanum). / Denz, Christopher R.; Zhang, Chi; Jia, Pingping; Du, Jianfeng; Huang, Xupei; Dube, Syamalima; Thomas, Anish; Poiesz, Bernard J.; Dube, Dipak K.

In: Cardiovascular Toxicology, Vol. 11, No. 3, 01.09.2011, p. 235-243.

Research output: Contribution to journalArticle

Denz, CR, Zhang, C, Jia, P, Du, J, Huang, X, Dube, S, Thomas, A, Poiesz, BJ & Dube, DK 2011, 'Absence of mutation at the 50-upstream promoter region of the TPM4 gene from cardiac mutant axolotl (Ambystoma mexicanum)', Cardiovascular Toxicology, vol. 11, no. 3, pp. 235-243. https://doi.org/10.1007/s12012-011-9117-z
Denz, Christopher R. ; Zhang, Chi ; Jia, Pingping ; Du, Jianfeng ; Huang, Xupei ; Dube, Syamalima ; Thomas, Anish ; Poiesz, Bernard J. ; Dube, Dipak K. / Absence of mutation at the 50-upstream promoter region of the TPM4 gene from cardiac mutant axolotl (Ambystoma mexicanum). In: Cardiovascular Toxicology. 2011 ; Vol. 11, No. 3. pp. 235-243.
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