Rapid decrease in thymidine kinase activity of mouse cell temperature-sensitive mutants at a non-permissive...

A rapid decrease in the incorporation of [3H]thymidine into DNA at a non-permissive temperature was observed in two temperature-sensitive mutants that were isolated from mouse FM3A cells. This change was not due to a decrease in the rate of DNA replication, but was closely associated with a decrease in thymidine kinase activity of these cells. Experiments to test thermolability of thymidine kinase in extracts showed that there are two components of the thymidine kinase, but there was no alteration in the sensitivity of the enzyme to high temperature. Also, the decrease in enzyme activity in the temperature-sensitive mutants at the non-permissive temperature occurred much faster than expected from the half-life of the enzyme in wild-type cells, which was measured in the presence of cycloheximide. These results suggested that the enzyme was somehow rapidly inactivated, or degraded, in the cells at the non-permissive temperature.