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Publication : Control of the murine phosphofructokinase-A gene during muscle differentiation.

First Author  Gekakis N Year  1994
Journal  Biochemistry Volume  33
Issue  7 Pages  1771-7
PubMed ID  8110779 Mgi Jnum  J:17108
Mgi Id  MGI:65161 Doi  10.1021/bi00173a021
Citation  Gekakis N, et al. (1994) Control of the murine phosphofructokinase-A gene during muscle differentiation. Biochemistry 33(7):1771-7
abstractText  The muscle-specific isoform of phosphofructokinase (PFK-A) is induced during muscle development. To understand expression of PFK at the molecular level, transcription of the mouse PFK-A gene was examined during C2 myoblast differentiation to myotubes. PFK-A gene transcription increased 5-7-fold during differentiation in vitro. To identify cis-acting elements which direct muscle-specific transcription of the PFK-A gene, its 5'-flanking region and first exon were cloned and characterized. S1 nuclease protection and primer extension assays showed four sites of transcription initiation at 106, 105, 88, and 87 bp upstream of the translation initiation codon. Stable transfection of fusion genes linking -1900 to +99 of PFK-A 5'-flanking sequence to chloramphenicol acetyltransferase coding sequences into myogenic C2 cells did not confer muscle-specific expression. However, larger fragments of PFK-A 5'-flanking region (-5800 to +99) showed muscle-specific expression by transient transfection assay. The sequences directing muscle-specific transcription were further defined by linking various PFK-A upstream fragments to the luciferase gene under the control of the PFK-A proximal promoter, -335 to +99 bp. We found DNA sequence responsible for muscle-specific expression of the PFK-A gene between -4800 and -3900 bp.
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