Regulation of the rat muscle glycogen phosphorylase-encoding gene during muscle cell development.
The muscle isozyme of glycogen phosphorylase (MGP) catalyzes the hydrolysis hydrolysis of intracellular glycogen in mammalian tissues and is produced in skeletal muscle, brain and heart. The MGP gene is developmentally and neutrally regulated in skeletal muscle, but little is known about the gene's transcriptional regulation. We have isolated and characterized the 5' flanking region of rat MGP. Truncated portions of the MGP 5' flanking region were coupled to the bacterial cat reporter gene and used in transient transfection assays in the mouse muscle C2C12 cell line. The region between -211 and +62 contained the smallest regulatory domain capable of demonstrating developmentally regulated myogenic expression in C2C12 cells. This was in contrast with findings from another investigation that transfected this cell line with human MGP [Lockyer and McCracken, J. Biol. Chem. 266 (1991) 20262-20269]. A 172-nucleotide (nt) region between -839 and -666 functioned as a potent enhancer in C2C12 cells when coupled to its cognate promoter, but not when coupled to a simian virus 40 promoter. This rat MGP enhancer region is 78% identical to a comparable region of the human MGP 5' flanking region, but contains only one putative regulatory element that has been previously identified in other muscle genes. These data suggest that rat MGP transcription in C2C12 muscle cells is modulated by a potent enhancer that utilizes novel regulatory elements.
Department of Neurology, University of California, Davis 95616-8647.
B E
BE
Froman
K R
KR
Herrick
F A
FA
Gorin
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D004742:Enhancer Elements, Genetic; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis*; D008099:Liver / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D024510:Muscle Development; D009132:Muscles / Q000166:cytology / Q000201:enzymology*; D006005:Phosphorylases / Q000096:biosynthesis*; D011401:Promoter Regions, Genetic; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013539:Simian virus 40 / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D014599:Uterus / Q000201:enzymology
D007527:Isoenzymes; D015500:Chloramphenicol O-Acetyltransferase; D006005:Phosphorylases