Cloning of cDNAs for fructose 6-phosphate 2-kinase/fructose 2,6-bisphosphatase from frog skeletal muscle and liver, and their expression in skeletal muscle.
Frog (Rana catesbeiana) skeletal muscle (M-type) and liver (L-type) cDNAs of fructose 6-phosphate 2-kinase/fructose 2,6-bisphosphatase were isolated from lambda gt10 phage cDNA library. The full-length L-type cDNA (1829 bp) encodes a 469 amino acids subunit (M(r) 54,800), while the M-type cDNA (1792 bp) encodes 455 amino acids (M(r) 52,901). The amino acid sequence of the M-type isozyme is identical to that of the L-type isozyme except for the N-terminus. The N-terminal 30 amino acids of the L-type isozyme are replaced by an unique sequence of 16 amino acids in the M-type isozyme. Both L- and M-type cDNAs were detected also in a lambda gt10 phage library of skeletal muscle. Relative amount of the M- and L-type mRNAs is skeletal muscle was determined by the reverse transcription-polymerase chain reaction method. The M/L mRNA ratio in frog skeletal muscle shows seasonal variations, being 0.56/1 in early summer and 5.3/1 in winter. These results suggest that there is a seasonal change in the isozyme composition and that the glycolysis in frog skeletal muscle may be regulated by type of the isozyme synthesized.
Sakai
A
A
Department of Chemistry, Osaka Medical College, Japan.
Watanabe
F
F
Furuya
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D015870:Gene Expression; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000235:genetics; D009132:Muscles / Q000201:enzymology*; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases / Q000096:biosynthesis* / Q000235:genetics; D010770:Phosphotransferases / Q000096:biosynthesis* / Q000235:genetics; D011061:Poly A / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D011892:Rana catesbeiana; D051381:Rats; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D007527:Isoenzymes; D009097:Multienzyme Complexes; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D010770:Phosphotransferases; D025481:Phosphofructokinase-2; D010744:Phosphoric Monoester Hydrolases