cDNA Cloning and Characterization of Murine Transcriptional Enhancer Factor-1-related Protein 1, a Transcription Factor That Binds to the M-CAT Motif (*)
- From the (1)Molecular Medicine Division, Beth Israel Hospital and Department of Medicine, Harvard Medical School, Boston, Massachusetts 02215 and
- (2)Department of Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia, South Carolina 29208
- ¶To whom correspondence should be addressed: Dept. of Developmental Biology and Anatomy, University of South Carolina, School of Medicine, Columbia, SC 29208. Tel.: 803-733-3215; Fax: 803-733-1533; :shimizu{at}dcsmserver.med.sc.edu.
Abstract
The M-CAT motif is a cis-regulatory DNA sequence that is essential for muscle-specific transcription of several genes. Previously,
we had shown that both muscle-specific (A1) and ubiquitous (A2) factors bind to an essential M-CAT motif in the myosin heavy
chain β gene and that the ubiquitous factor is transcriptional enhancer factor (TEF)-1. Here we report the isolation of mouse
cDNAs encoding two forms (a and b) of a TEF-1-related protein, TEFR1. The TEFR1a cDNA encodes a 427-amino acid protein. The
coding region of TEFR1b is identical to 1a in both nucleotide and predicted amino acid sequence except for the absence of
43 amino acids downstream of the TEA DNA-binding domain. Three TEFR1 transcripts (
7, 
3.5, and 
2 kilobase pairs) are enriched in differentiated skeletal muscle (myotubes) relative to undifferentiated skeletal muscle (myoblasts)
and non-muscle cells in culture. In situ hybridization analysis indicated that TEFR1 transcripts are enriched in the skeletal muscle lineage during mouse embryogenesis.
Transient expression of fusion proteins of TEFR1 and the yeast GAL4 DNA-binding domain in cell lines activated the expression
of chloramphenicol acetyltransferase (CAT) reporter constructs containing GAL4 binding sites, indicating that TEFR1 contains
an activation domain. An anti-TEFR1 polyclonal antibody supershifted the muscle-specific M-CAT•A1 factor complex in gel mobility
shift assays, suggesting that TEFR1 is a major component of this complex. Our results suggest that TEFR1 might play a role
in the embryonic development of skeletal muscle in the mouse.
Footnotes
-
↵* This work was supported in part by National Institutes of Health Grant AR41893 and the American Heart Association, Massachusetts Affiliate, Grant 13-519-912. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank(™)/EMBL Data Bank with accession number(s) L26343[GenBank] (TEFR1a) and L26344[GenBank] (TEFR1b).
-
↵1 The abbreviations used are:
- TEF-1
-
transcriptional enhancer factor-1
- CAT
-
chloramphenicol acetyltransferase
- aa
-
amino acid(s)
- ETF
-
embryonic TEA domain-containing factor
- GST
-
glutathione S-transferase
- nt
-
nucleotide(s)
- PAGE
-
polyacrylamide gel electrophoresis
- RT
-
reverse transcription
- PCR
-
polymerase chain reaction
- mTEF1
-
mouse TEF-1
- cTEF1
-
chicken TEF-1
- TEFR1
-
TEF-1-related protein 1
- bp
-
base pair(s)
- kb
-
kilobase pair(s).
-
- Received August 16, 1995.
- Revision received October 16, 1995.
- © 1996 by The American Society for Biochemistry and Molecular Biology, Inc.
