cDNA sequence of rat liver fructose-1,6-bisphosphatase and evidence for down-regulation of its mRNA by insulin.
A coding-length clone of rat liver fructose-1,6-bisphosphatase (EC 3.1.3.11) was isolated by immunological screening of a cDNA library in lambda gt11. Its identity was verified by comparing the deduced amino acid sequence with that obtained by direct sequencing of a complete set of CNBr and proteolytic peptides from the purified protein. The enzyme subunit is composed of 362 amino acids and has N-acetylvaline as the amino-terminal residue. The cDNA, 1255 base pairs (bp) long, consisted of 1086 bp of coding region, 15 bp of 5' untranslated sequence, and 154 bp at the 3' untranslated end. The 3' untranslated sequence contained a polyadenylylation signal (AATAAA) followed after 30 bp by a stretch of 7 adenines at the end of the clone. The deduced amino acid sequence was identical to the primary sequence of the protein and confirmed the alignment of five nonoverlapping peptides. It also confirmed the 27-residue extension, unique to the rat liver subunit, ending with a carboxyl-terminal phenylalanine. RNA blot analyses using the radiolabeled liver cDNA as a probe revealed a single band of fructose-1,6-bisphosphatase mRNA, 1.4 kilobases long, in liver and kidney but not in nongluconeogenic tissues. Fructose-1,6-bisphosphatase mRNA was increased 10-fold in livers from diabetic rats and was reduced to control levels after 24 hr of insulin treatment, suggesting that the changes in enzyme activity observed in diabetes and after insulin treatment are due to alterations in mRNA abundance.
el-Maghrabi
M R
MR
Department of Physiology and Biophysics, State University of New York, Stony Brook 11794.
Pilkis
J
J
Marker
A J
AJ
Colosia
A D
AD
D'Angelo
G
G
Fraser
B A
BA
Pilkis
S J
SJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003062:Codon; D004247:DNA / Q000187:drug effects / Q000235:genetics*; D006597:Fructose-Bisphosphatase / Q000235:genetics*; D007328:Insulin / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics*; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D003062:Codon; D007328:Insulin; D012333:RNA, Messenger; D004247:DNA; D006597:Fructose-Bisphosphatase