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PMID: 8114720 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A novel myogenic regulatory circuit controls slow/cardiac troponin C gene transcription in skeletal muscle.

Molecular and cellular biology ·Vol. 14 ·No. 3 ·1994-03-00 ·Pages 1870-85

Parmacek MS, Ip HS, Jung F, Shen T, Martin JF, Vora AJ, Olson EN, Leiden JM

Abstract

The slow/cardiac troponin C (cTnC) gene is expressed in three distinct striated muscle lineages: cardiac myocytes, embryonic fast skeletal myotubes, and adult slow skeletal myocytes. We have reported previously that cTnC gene expression in cardiac muscle is regulated by a cardiac-specific promoter/enhancer located in the 5' flanking region of the gene (bp -124 to +1). In this report, we demonstrate that the cTnC gene contains a second distinct and independent transcriptional enhancer which is located in the first intron. This second enhancer is skeletal myotube specific and is developmentally up-regulated during the differentiation of myoblasts to myotubes. This enhancer contains three functionally important nuclear protein binding sites: a CACCC box, a MEF-2 binding site, and a previously undescribed nuclear protein binding site, designated MEF-3, which is also present in a large number of skeletal muscle-specific transcriptional enhancers. Unlike most skeletal muscle-specific transcriptional regulatory elements, the cTnC enhancer does not contain a consensus binding site (CANNTG) for the basic helix-loop-helix (bHLH) family of transcription factors and does not directly bind MyoD-E12 protein complexes. Despite these findings, the cTnC enhancer can be transactivated by overexpression of the myogenic bHLH proteins, MyoD and myogenin, in C3H10T1/2 (10T1/2) cells. Electrophoretic mobility shift assays demonstrated changes in the patterns of MEF-2, CACCC, and MEF-3 DNA binding activities following the conversion of 10T1/2 cells into myoblasts and myotubes by stable transfection with a MyoD expression vector. In particular, MEF-2 binding activity was up-regulated in 10T1/2 cells stably transfected with a MyoD expression vector only after these cells fused and differentiated into skeletal myotubes. Taken together, these results demonstrated that distinct lineage-specific transcriptional regulatory elements control the expression of a single myofibrillar protein gene in fast skeletal and cardiac muscle. In addition, they show that bHLH transcription factors can indirectly transactivate the expression of some muscle-specific genes.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Cell Line Consensus Sequence DNA-Binding Proteins/chemistry,metabolism Enhancer Elements, Genetic Gene Expression Regulation Helix-Loop-Helix Motifs Introns Macromolecular Substances Mice Molecular Sequence Data Muscles/physiology MyoD Protein/metabolism Myogenin/metabolism Nuclear Proteins/metabolism Oligodeoxyribonucleotides/chemistry RNA, Messenger/genetics Sequence Alignment Sequence Homology, Amino Acid Sequence Homology, Nucleic Acid Transcription, Genetic Transcriptional Activation Troponin/genetics Troponin C
Chemicals
DNA-Binding Proteins Macromolecular Substances MyoD Protein Myog protein, mouse Myogenin Nuclear Proteins Oligodeoxyribonucleotides RNA, Messenger Troponin Troponin C
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Parmacek M S
Department of Medicine, University of Chicago, Illinois 60637.
Ip H S
Jung F
Shen T
Martin J F
Vora A J
Olson E N
Leiden J M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.3.1870-1885.1994
Published
1994-03-00
Pages
1870-85
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358545
Subset
IM
Grants
NHLBI NIH HHS · 1RO1HL51145-01 · United States
External Links
PubMed source DOI: 10.1128/mcb.14.3.1870-1885.1994 PMC Full Text View journal details
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