Determination of isozyme-specific mRNAs of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in glutamate obese rats by RNA probes.
To study the possible alterations in the metabolism of fructose 2,6-bisphosphate in glutamate induced obese rats, the mRNA levels of different isozymes coding for the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/F2,6Pase) were determined in various rat tissues obtained from fed and starved animals. For Northern analysis and dot blotting radioactively labeled RNA probes were synthesized by in vitro transcription. A modified ligase-free subcloning method was applied to obtain the plasmid vectors containing specific fragments of the cDNAs coding for the muscle, liver and heart isozymes of PFK-2/F2,6Pase. In liver but not in heart and skeletal muscle of glutamate obese rats the content of mRNA of PFK-2/F2,6Pase is significantly lower than in normal rats. As in normal rats short-term starvation has no effect on mRNA levels of the bifunctional enzyme. These results are compared with the maximum activities of PFK-2 and F2,6Pase determined in cell-free extracts prepared from the three organs.
Temesgen
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Institut für Biochemie, Bereich Medizin, Universität, Leipzig, Germany.
Eschrich
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K
D000818:Animals; D016375:Antisense Elements (Genetics); D015152:Blotting, Northern; D005971:Glutamates; D018698:Glutamic Acid; D007527:Isoenzymes / Q000096:biosynthesis*; D008099:Liver / Q000201:enzymology; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009765:Obesity / Q000139:chemically induced / Q000201:enzymology*; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases / Q000096:biosynthesis*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis* / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D013217:Starvation / Q000201:enzymology; D014158:Transcription, Genetic
D016375:Antisense Elements (Genetics); D005971:Glutamates; D007527:Isoenzymes; D015347:RNA Probes; D012333:RNA, Messenger; D018698:Glutamic Acid; D017853:Phosphotransferases (Alcohol Group Acceptor); D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases