Increased translatable messenger ribonucleic acid for argininosuccinate synthetase in canavanine-resistant human cells.
The level of argininosuccinate synthetase activity in the human tissue culture cell line RPMI 2650 was 6-fold higher when citrulline was substituted for arginine in the culture medium. Canavanine-resistant (Canr) variants were isolated and had enzyme activity up to 25 nmol min-1 (mg of protein)-1 or 180-fold higher than that of the wild-type cells grown in arginine. The differences in enzyme activity were paralleled by differences in the amount of enzyme determined immunologically. The micrograms of enzyme per milligrams of protein, determined by complement fixation, were 0.03 for wild-type cells grown in arginine, 0.29 for wild-type cells grown in citrulline, and 6.73 for a Canr variant. In vivo labeling studies suggested increased synthesis of argininosuccinate synthetase in Canr cells, and in vitro translation of poly(adenylic acid) [poly(A)] messenger ribonucleic acid (mRNA) from wild-type and Canr cells confirmed a quantitatively compatible increase in translatable poly(A) mRNA for the enzyme in Canr cells. No precursor for the enzyme was recognized by using in vitro translation, and the poly(A) mRNA for the enzyme had a sedimentation value of 16 S by sucrose-gradient analysis. The levels of argininosuccinate synthetase activity in the Canr cells were similar to those found in normal liver.
D000818:Animals; D001124:Argininosuccinate Synthase / Q000235:genetics*; D002172:Canavanine / Q000494:pharmacology*; D002460:Cell Line; D004351:Drug Resistance; D014644:Genetic Variation; D006801:Humans; D007118:Immunoassay; D007951:Leukemia, Myeloid; D008025:Ligases / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism
D012333:RNA, Messenger; D002172:Canavanine; D008025:Ligases; D001124:Argininosuccinate Synthase