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Inhibition of Thymidine Phosphorylation and DNA and Histone Synthesis in Ehrlich Ascites Carcinoma¹

Departments of Biochemistry and Medicinal Chemistry, University of North Carolina, Chapel Hill, North Carolina 27514

The time course of phosphorylation of thymidine and the effects of various inhibitors on the phosphorylation of thymidine and on DNA synthesis in Ehrlich ascites carcinoma cells in vitro were studied with a double-isotope labeling method. Hydroxyurea, mitomycin C, cycloheximide, puromycin, 1-ß-D-arabinofuranosylcytosine, 5-fluoroorotic aldehyde, and the thiosemicarbazones of 2-phthalimido aliphatic aldehydes inhibit incorporation of thymidine-2-¹⁴C into DNA and produce an accumulation of label in thymidine 5'-triphosphate in Ehrlich ascites carcinoma cells in vitro.

A concentration (0.01 mM) of 1-ß-D-arabinofuranosyl-cytosine that inhibits incorporation of thymidine into DNA by 75% also depresses incorporation of labeled glycine into slightly lysine-rich histones (70%), lysine-rich histones (30%), and arginine-rich histones (20%); incorporation of labeled glycine into total cytoplasmic proteins is not inhibited by this concentration of 1-ß-D-arabinofuranosylcytosine in short-term experiments.

¹ Aided by American Cancer Society Grants E-24J and In-15-?? and NIH Grants GM-08318-08 and CA-10729.

Received 11/13/69. Accepted 7/ 2/70.