Phosphofructokinase isozyme expression during myoblast differentiation.
Isozyme expression of phosphofructokinase (PFK), the key regulatory enzyme for glycolysis, was studied during differentiation of mouse C2 myoblasts to myotubes. The total PFK activity increased 20-fold during in vitro myogenesis. The rate of synthesis, relative to the rate of total protein synthesis, measured by pulse labeling and immunoprecipitation was lowest for muscle PFK (PFK-A), 0.008% in myoblasts, while those for liver (PFK-B) and brain (PFK-C) PFK were 0.017 and 0.014%, respectively. The relative rate of PFK-A synthesis increased sharply (5-fold) at an initial period of differentiation (8 h) and reached maximum of 10-fold at 48 h, to make PFK-A the major isoform synthesized in myotubes. The relative rates of synthesis for both PFK-B and PFK-C did not change drastically, decreasing slightly at 8 h, but were restored to 1.5-2-fold of myoblasts. cDNA sequences coding for mouse muscle PFK were cloned and used along with those for mouse liver PFK, which we have previously cloned, to measure by Northern blot analysis under highly stringent conditions the steady-state mRNA concentrations for muscle and liver PFK during C2 differentiation. The hybridizable mRNA level for PFK-A increased gradually, reaching 13-fold at 48 h when 80% of cells was fused to myotubes. The PFK-A mRNA level at 96 h was 90-fold of that for myoblasts. In contrast, the mRNA level for PFK-B increased slightly during differentiation, showing a maximum of 4-fold at 96 h. These results indicate isozyme-specific control of muscle PFK gene expression during C2 myoblast differentiation.
Gekakis
N
N
Department of Nutrition, Harvard School of Public Health, Boston, Massachusetts 02115.
Gehnrich
S C
SC
Sul
H S
HS
D000818:Animals; D001921:Brain / Q000201:enzymology; D002454:Cell Differentiation; D002460:Cell Line; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D006019:Glycolysis; D007527:Isoenzymes / Q000378:metabolism*; D008099:Liver / Q000201:enzymology; D051379:Mice; D009132:Muscles / Q000166:cytology / Q000201:enzymology*; D010732:Phosphofructokinase-1 / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics
D007527:Isoenzymes; D012333:RNA, Messenger; D004247:DNA; D010732:Phosphofructokinase-1