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PMID: 1582413 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Co-operativity of functional domains in the muscle-specific transcription factor Myf-5.

The EMBO journal ·Vol. 11 ·No. 5 ·1992-05-00 ·Pages 1843-55

Winter B, Braun T, Arnold HH

Abstract

Myf-5 is a member of a family of muscle-specific transcription factors that activate myogenesis in 10T1/2 fibroblasts. Here we report the analysis of Myf-5 structural domains that are responsible for its biological activity. Site-directed mutagenesis revealed that two clusters of basic amino acids within a conserved basic region and two amphipathic helices within the adjacent HLH domain are essential for sequence-specific DNA binding and hetero-oligomerization, respectively. Transcriptional activation by Myf-5 requires two additional domains located in the amino- and carboxyl-termini. The two domains apparently co-operate since deletion of either one results in inactivation. Chimeric proteins between DNA binding domain of the yeast transcription factor GAL4 and the separate Myf-5 transactivator domains exhibit activity that is enhanced when both regions are combined. Dimerization of Myf-5 with the ubiquitously expressed bHLH protein E12 not only increases the affinity for DNA but also stimulates transactivation independently of DNA binding. The Myf-5 transactivator domains are dependent for activity on a specific amino acid sequence motif within the basic region when Myf-5 activity is mediated through the E-box DNA recognition sequence but not when DNA binding occurs through the GAL4 DNA binding domain. This demonstrates that muscle-specific transactivation by Myf-5 requires the collaboration of two activation domains and the DNA binding region in addition to sequence-specific DNA binding. Transcriptional activation and interaction with DNA are executed by separable domains; however, transactivation is influenced by the basic region in a manner distinguishable from DNA binding.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cells, Cultured DNA/genetics DNA-Binding Proteins/metabolism Fungal Proteins/metabolism Mice Molecular Sequence Data Muscle Proteins/genetics,metabolism Muscles/metabolism Mutagenesis, Site-Directed Myogenic Regulatory Factor 5 Recombinant Fusion Proteins/genetics,metabolism Saccharomyces cerevisiae Proteins Trans-Activators Transcription Factors/genetics,metabolism Transcription, Genetic Transcriptional Activation
Chemicals
DNA-Binding Proteins Fungal Proteins GAL4 protein, S cerevisiae Muscle Proteins Myf5 protein, mouse Myogenic Regulatory Factor 5 Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Trans-Activators Transcription Factors DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Winter B
Department of Toxicology, University of Hamburg, Medical School, FRG.
Braun T
Arnold H H
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-05-00
Pages
1843-55
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556642
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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