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Molecular and Cellular Biology, January 1999, p. 296-306, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Poly(ADP-Ribose) Polymerase Binds with
Transcription Enhancer Factor 1 to MCAT1 Elements To Regulate
Muscle-Specific Transcription
Alison J.
Butler and
Charles P.
Ordahl*
Department of Anatomy and Cardiovascular
Research Institute, University of California San Francisco, San
Francisco, California 94143-0452
Received 27 May 1998/Returned for modification 10 July
1998/Accepted 2 October 1998
Striated muscle-specific expression of the cardiac troponin T
(cTNT) gene is mediated through two MCAT elements that act via binding
of transcription enhancer factor 1 (TEF-1) to the MCAT core motifs and
binding of an auxiliary protein to nucleotides flanking the 5' side of
the core motif. Using DNA-protein and protein-protein binding
experiments, we identified a 140-kDa polypeptide that bound both the
muscle-specific flanking sequences of the most distal MCAT1 element and
TEF-1. Screening of an expression library with the MCAT1 element
yielded a cDNA encoding a truncated form of poly(ADP-ribose) polymerase
(PARP). Endogenous PARP from embryonic tissue nuclear extracts migrated
as a 140-kDa protein. Recombinant full-length PARP preferentially bound
the wild-type MCAT1 element and was shown to physically interact with
TEF-1. In addition, endogenous TEF-1 could be coimmunoprecipitated with PARP from extracts of primary skeletal muscle cells. Recombinant PARP
was able to ADP-ribosylate TEF-1 in vitro. Inhibition of the enzymatic
activity of PARP repressed expression of an MCAT1-dependent reporter in
transiently transfected primary muscle cells. Together, these data
implicate PARP as the auxiliary protein that binds with TEF-1 to the
MCAT1 element to provide muscle-specific gene transcription.
*
Corresponding author. Mailing address: Department of
Anatomy and Cardiovascular Research Institute, Box 0452, University of California San Francisco, 513 Parnassus Ave., San Francisco, CA 94143. Phone: (415) 476-4051. Fax: (415) 476-4845. E-mail:
ordahl{at}itsa.ucsf.edu.
Molecular and Cellular Biology, January 1999, p. 296-306, Vol. 19, No. 1
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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