Subject(s)
Amino Acid Oxidoreductases/metabolism , Aminoisobutyric Acids/metabolism , Tetrahymena pyriformis/enzymology , Amino Acid Oxidoreductases/isolation & purification , Animals , Cations, Divalent/pharmacology , Detergents/pharmacology , Flavin-Adenine Dinucleotide/pharmacology , Hydrogen-Ion Concentration , Kinetics , TemperatureABSTRACT
The operation of reductive pyrimidine catabolic and reutilization pathway in Tetrahymena pyriformis was investigated. Consistent with the proposed catabolic interconversions, radioactivity from [2,6-14C2]thymidine was recovered in respired CO2 30 min after its addition to the culture, whereas, consistent with the proposed anabolic interconversions, over 50% of the incorporated label was recovered in cellular macromolecules other than DNA 12 h after its addition. The chromatographic recovery of 14C radioactivity in monosaccharides from [2,6-14C2]thymidine as well as from [2-14C]methylmalonic acid, a key reutilization intermediate in this proposed pathway, further substantiated the operation of the required anabolic interconversions in this organism.
Subject(s)
Malonates/metabolism , Methylmalonic Acid/metabolism , Tetrahymena pyriformis/metabolism , Thymidine/metabolism , Animals , Carbohydrates/biosynthesis , Carbon Radioisotopes , DNA/biosynthesis , Kinetics , Lipids/biosynthesis , Protein Biosynthesis , RNA/biosynthesis , Radioisotope Dilution TechniqueABSTRACT
By phenol extraction and DEAE cellulose column chromatography, tRNA was isolated from Tetrahymena pyriformis strain GL. Following acid hydrolysis of the tRNA, the methylated purine content was determined by Dowex 50 column chromatography and paper chromatography. The most abundant methylated guanine derivative was found to be N2-DMG. Also present were 1-MG, N2-MG, and 7-MG. The most abundant methylated adenine was found to be 1-MA; no 2-MA was detected. Small amounts of the N6-methyladenines were detected.
Subject(s)
Guanine/analysis , RNA, Transfer/analysis , Tetrahymena pyriformis/analysis , Adenine/analogs & derivatives , Adenine/analysis , Animals , Chromatography , Guanine/analogs & derivatives , Methylation , PurinesABSTRACT
Growth (dry weight accumulation) of Sordaria fimicola in standing liquid culture (sucrose-nitrate-salts-vitamins) is inhibited by the presence of 5 muM 5-fluorouracil in the medium. This inhibition is completely prevented by uracil, deoxyuridine, and 5-bromouracil, partly prevented (40 to 90% of growth observed without 5-fluorouracil) by uridine, thymidine, and 5-bromodeoxyuridine, and slightly prevented by trifluorothymine, cytosine, cytidine, deoxycytidine, and 5-methylcytosine (all at 0.5 to 1 mM). Thymidine and thymine riboside were without any apparent effect. Growth is also inhibited by 0.2 mM 6-azauracil, and this inhibition was completely prevented by uracil and uridine, partly prevented by deoxyuridine, 5-bromouracil, cytidine, and 5-methylcytosine, and slightly prevented by thymine, thymidine, 5-bromodeoxyuridine, cytosine, and deoxycytidine. The data suggest that the observed inhibition of growth by 5-fluorouracil is due to inhibition of both ribonucleic acid and deoxyribonucleic acid synthesis. The data also allow inferences concerning pyrimidine interconversions in S. fimicola; i.e., thymine can be anabolized to thymidylic acid without first being demethylated, although demethylation appears to occur also.
