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Molecular and Cellular Biology, August 2003, p. 5143-5164, Vol. 23, No. 15
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.15.5143-5164.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Transcription Enhancer Factor 1 Binds Multiple Muscle MEF2 and A/T-Rich Elements during Fast-to-Slow Skeletal Muscle Fiber Type Transitions

Natalia Karasseva,1 Gretchen Tsika,2 Juan Ji,2 Aijing Zhang,1 Xiaoqing Mao,1 and Richard Tsika1,2,3*

Department of Biochemistry, School of Medicine,1 Department of Biomedical Sciences, School of Veterinary Medicine,2 Dalton Cardiovascular Research Center, University of Missouri—Columbia, Columbia, Missouri 652113

Received 17 March 2003/ Returned for modification 29 April 2003/ Accepted 13 May 2003

In adult mouse skeletal muscle, ß-myosin heavy chain (ßMyHC) gene expression is primarily restricted to slow type I fibers; however, its expression can be induced in fast type II fibers in response to a sustained increase in load-bearing work (mechanical overload [MOV]). Our previous ßMyHC transgenic and protein-DNA interaction studies have identified an A/T-rich element (ßA/T-rich -269/-258) that is required for slow muscle expression and which potentiates MOV responsiveness of a 293-bp ßMyHC promoter (ß293wt). Despite the GATA/MEF2-like homology of this element, we found binding of two unknown proteins that were antigenically distinct from GATA and MEF2 isoforms. By using the ßA/T-rich element as bait in a yeast one-hybrid screen of an MOV-plantaris cDNA library, we identified nominal transcription enhancer factor 1 (NTEF-1) as the specific ßA/T-rich binding factor. Electrophoretic mobility shift assay analysis confirmed that NTEF-1 represents the enriched binding activity obtained only when the ßA/T-rich element is reacted with MOV-plantaris nuclear extract. Moreover, we show that TEF proteins bind MEF2 elements located in the control region of a select set of muscle genes. In transient-coexpression assays using mouse C2C12 myotubes, TEF proteins transcriptionally activated a 293-bp ßMyHC promoter devoid of any muscle CAT (MCAT) sites, as well as a minimal thymidine kinase promoter-luciferase reporter gene driven by three tandem copies of the desmin MEF2 or palindromic Mt elements or four tandem ßA/T-rich elements. These novel findings suggest that in addition to exerting a regulatory effect by binding MCAT elements, TEF proteins likely contribute to regulation of skeletal, cardiac, and smooth muscle gene networks by binding select A/T-rich and MEF2 elements under basal and hypertrophic conditions.

* Corresponding author. Mailing address: Department of Biochemistry, E102 Vet. Med. Bldg., University of Missouri—Columbia, 1600 Rollins Rd., Columbia, MO 65211. Phone: (573) 884-4547. Fax: (573) 884-6890. E-mail: tsikar{at}missouri.edu.

Molecular and Cellular Biology, August 2003, p. 5143-5164, Vol. 23, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.15.5143-5164.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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