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. 1990 Aug;87(15):5623–5627. doi: 10.1073/pnas.87.15.5623

MyoD binds cooperatively to two sites in a target enhancer sequence: occupancy of two sites is required for activation.

H Weintraub 1, R Davis 1, D Lockshon 1, A Lassar 1
PMCID: PMC54379  PMID: 2377600

Abstract

MyoD is a master regulatory gene for myogenesis. Its product, the MyoD protein, appears to act by binding to muscle-specific enhancer sequences. We show that MyoD binds cooperatively to two sites in the muscle-specific creatine kinase enhancer; this is dramatically reflected in dissociation-rate measurements. A deletion of the acidic N terminus (residues 3-56) results in a protein that binds normally to single sites but fails to bind cooperatively to two adjacent sites, suggesting a role of the N terminus in cooperative interactions. In transfection assays, a reporter gene flanked by a single MyoD binding site fails to be activated by cotransfected MyoD expression vectors. In contrast, a reporter with two or more MyoD binding sites is activated by wild-type MyoD but not by N-terminally deleted MyoD. A reporter gene with a single binding site, although not activated by MyoD, can nonetheless compete for expression with a reporter gene containing three sites. Thus, in vivo, a single site can bind MyoD, but occupancy of two or more sites is required for subsequent transcriptional activation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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