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Overexpression of Id protein inhibits the muscle differentiation program: in vivo association of Id with E2A proteins
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The helix-loop-helix (HLH) protein Id lacks the basic DNA-binding domain common to this class of proteins. In vitro experiments suggested that Id could associate tightly with two other HLH proteins encoded by the E2A gene, E12 and E47 (referred to here collectively as E proteins) and prevent their binding to a sequence present in the muscle creatine kinase (MCK) enhancer either as homo-oligomers or hetero-oligomers with MyoD. In this report we present evidence for the in vivo roles of Id and E proteins: (1) Id and E proteins co-fractionate and co-immunoprecipitate in whole-cell extracts prepared from myoblasts; (2) the loss of Id protein observed during the conversion of proliferating myoblasts into mature myotubes correlates with the formation of MyoD/E hetero-oligomeric complexes in whole-cell extracts (these complexes do not form when purified Id protein is added to the extracts); and (3) stable overexpression of Id mRNA and protein in the C2C12 muscle cell line inhibits differentiation in these cells 16 hr post-induction. The myotubes that do eventually form 48 hr post-induction have no detectable Id protein in the nucleus despite the persistence of exogenous Id mRNA. These data support a model in which Id can inhibit muscle cell differentiation by associating with E proteins and preventing them from forming active hetero-oligomeric complexes with the muscle determination gene products.
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“When comparing Pax7+ cells in transcriptional cluster 4 with Pax7− cells in the same cluster, we found that Pax7+ cells showed significant upregulation of Id1, Id2 and Id3, which inhibit differentiation (Jen et al., 1992), and of the cell-cycle gene Ccnd1 (Fig. S3D).”
From Aging induces aberrant state transition kinetics in murine muscle stem cells (2020) · cites this paper as Jen et al., 1992
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Metadata
- Authors
- Jen, Y., Weintraub, H., & Benezra, R.
- Affiliations
- Memorial Sloan Kettering Cancer Center (US)
- ORCID
- Benezra, R
- Venue
- Genes & Development
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 1992-08
- Volume
- 6
- Issue
- 8
- Pages
- 1466-1479
- ISSN
- 0890-9369 (ISSN-L)
- 1549-5477
- OpenAlex
- W2101927529
- Cited by
- 471
- References
- 47
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 5 · 2023: 1 · 2022: 5 · 2021: 10 · 2020: 9 · 2019: 7 · 2018: 9 · 2017: 9 · 2016: 6 · 2015: 7 · 2014: 15 · 2013: 4 · 2012: 8
- Type
- posted content
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Genes & Development (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Howard Hughes Medical Institute
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- Id proteins, E12, E47, helix-loop-helix proteins, myogenesis, MyoD, hetero-oligomeric complexes, C2C12 myoblasts, transcriptional repression, myotubes
- MeSH terms
- Animals, Cell Differentiation, Cell Line, DNA-Binding Proteins, Fluorescent Antibody Technique, Muscles, Precipitin Tests, Repressor Proteins, Transcription Factors, Blotting, Northern, Inhibitor of Differentiation Protein 1, TCF Transcription Factors, Transcription Factor 7-Like 1 Protein
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