Expression of nuclear genes encoding the urea cycle enzymes, carbamoyl-phosphate synthetase I and ornithine carbamoyl transferase, in rat liver and intestinal mucosa.
RNA dot-blot, quantitative electron microscope immunocytochemistry, and electrophoretic immunoblotting techniques were employed to investigate the expression of carbamoyl-phosphate synthetase I (CPS) and ornithine carbamoyl transferase (OCT) genes in rat liver and intestinal mucosa. Comparing only those cell types in the two tissues which express these enzymes, we show that the concentration of CPS and OCT in hepatocyte mitochondria is 2.3-times and 1.2-times greater, respectively, than in intestinal epithelial cell mitochondria. As a percentage of total tissue protein, however, liver homogenates contain 10-20 times more CPS and 5-10 times more OCT than is found in intestinal mucosa. These relatively large differences in enzyme protein levels between the two tissues are not reflected by differences in their mRNA levels. As a percentage of total translational activity in vitro (based on incorporation of [35S]methionine), total liver mRNA directed synthesis of about twice as much precursor CPS (pCPS) and precursor OCT (pOCT) than did equivalent amounts of mRNA from intestinal mucosa. The ratio of pCPS and pOCT mRNA levels between the two tissues (2:1, liver:intestinal mucosa) was confirmed by dot-blot and Northern hybridizations employing specific cDNA probes. The sizes of the respective mRNAs were the same for the two tissues: about 6000 residues for pCPS mRNA and about 1700 residues for pOCT mRNA.
Ryall
J
J
Nguyen
M
M
Bendayan
M
M
Shore
G C
GC
D000818:Animals; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000235:genetics*; D002474:Cell-Free System; D004247:DNA; D006651:Histocytochemistry; D007120:Immunochemistry; D007413:Intestinal Mucosa / Q000201:enzymology*; D008025:Ligases / Q000235:genetics*; D008099:Liver / Q000201:enzymology*; D009693:Nucleic Acid Hybridization; D009954:Ornithine Carbamoyltransferase / Q000235:genetics*; D011061:Poly A; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D012156:Reticulocytes / Q000378:metabolism; D014508:Urea / Q000096:biosynthesis
D012333:RNA, Messenger; D011061:Poly A; D014508:Urea; D004247:DNA; D009954:Ornithine Carbamoyltransferase; D008025:Ligases; D002222:Carbamoyl-Phosphate Synthase (Ammonia)