Cell
Volume 66, Issue 2, 26 July 1991, Pages 305-315
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Article
Functional activity of myogenic HLH proteins requires hetero-oligomerization with E12/E47-like proteins in vivo

https://doi.org/10.1016/0092-8674(91)90620-EGet rights and content

Abstract

In this report we provide four lines of evidence indicating that E12/E47-like proteins interact in vivo with the myogenic HLH proteins MyoD and myogenin. First, cotransfection of MyoD and E47 in COS cells indicates that these factors synergistically enhance transcription of a reporter gene containing an oligomerized MyoD-binding site. Second, mobility-shift assays of muscle cell nuclear extracts, “double shifted” with specific antisera, have identified complexes binding to the MEF1 site that contain either MyoD or myogenin in association with El 2/E47-like proteins. Third, association with E47 alters the phosphorylation state of MyoD. Fourth, C3H10T1/2 cells expressing antisense E2A transcripts contain low levels of E2A gene products and display less terminal muscle differentiation when infected with retroviral MyoD or when challenged to differentiate with 5-azacytidine treatment. In addition we demonstrate that MyoD, in conjunction with E12/E47-like proteins, is functioning as a regulatory nodal point for activation of several other downstream muscle regulators.

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    Present address: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.

    #

    Present address: Department of Biology, University of California at San Diego, San Diego, California 92093.

    ∗∗

    Present address: The Rockefeller University, 1230 York Avenue, New York, New York 10021.

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