Inhibition of protein and nucleic acid synthesis of animal cells in vitro mediated by high molecular weight inhibitors in human liver extract.

1. The addition of human liver extract to HeLa cells induces a reversible inhibition of the incorporation of [3H] thymidine into the DNA, [3H] uridine into the RNA, and 14C-labelled amino acids into the protein of HeLa cells. The inhibitory effects appear after treatment for 1 h and reach a maximum after 4-8 h. These effects do not depend on a defective precursor penetration, isotopic dilution or degradation of labelled precursor (thymidine-degrading enzymes were inactivated by the addition of unlabelled thymine), reduced activity of thymidine and uridine kinase, medium impairment, or an impairment of the cell-membrane function. 2. The nucleic acid synthesis-inhibiting activity of the extract seems to be dependent on cellular protein synthesis but independent of RNA synthesis which indicates that the inhibitors act in an indirect way. Furthermore, the inhibitors seem to lack the tissue-specific character of chalones. 3. The extract contains separate inhibitors of DNA, RNA and protein synthesis. These inhibitors were found to have different physical-chemical characteristics and to be macromolecules with a protein or conjugated protein character (mol. wt. approx. 90 000). 4. The possibility that the activity of the high molecular weight inhibitors resides in low molecular weight factors (bound to protein carriers) was tested: No true low molecular weight inhibitors could be liberated by extraction with trichloroacetic acid/organic solvents or by dialysis/enzymatic treatments. Nucleosides such as thymidine, uridine, and cytidine, however, were liberated and could be shown to interfere with the uptake of [3H] thymidine/[3H] uridine.