Molecular cloning of the cDNA coding for regucalcin and its mRNA expression in mouse liver: the expression is stimulated by calcium administration.
The molecular cloning of the cDNA coding for a Ca2+-binding protein regucalcin and its mRNA expression in mouse liver were investigated. The cDNA clone encoding a regucalcin was isolated from a mouse liver cDNA library and sequenced. Analysis of the sequence of the cloned cDNA showed that the cDNA encoded the complete amino acid sequence of the mouse regucalcin molecule; the cDNA had an open reading frame of 897 bp. Mouse regucalcin was composed of 299 amino acid residues, and its molecular weight was estimated to be 33,406 Da. The amino acid sequence of mouse regucalcin had 94% homology, as compared with that of rat regucalcin. Northern blot analysis with the mouse liver cDNA probe revealed that mouse regucalcin mRNA was mainly present in the liver but only slightly in the kidney with a size of 1.8 kb. Hepatic regucalcin mRNA level of male mouse was higher than that of female mouse. A single intraperitoneal administration of calcium chloride (5, 15, and 30 mg Ca2+/100 g body weight) to mice induced a remarkable increase in regucalcin mRNA in the liver; the increase in regucalcin mRNA levels at 30 min after calcium administration was dose-dependent. The present results demonstrate that regucalcin mRNA in mice is uniquely expressed in the liver, and that its expression is stimulated by calcium administration.
Murata
T
T
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Shizuoka City, Japan.
Yamaguchi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002118:Calcium / Q000008:administration & dosage / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000235:genetics*; D002265:Carboxylic Ester Hydrolases; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005260:Female; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis / Q000187:drug effects / Q000235:genetics*; D017422:Sequence Analysis, DNA; D015238:Sulfotransferases; D014018:Tissue Distribution
D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; C513324:Rgn protein, mouse; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; D002265:Carboxylic Ester Hydrolases; C513325:Rgn protein, rat; D002118:Calcium