Stimulation of thymidine kinase activity by chloramphenicol in Naegleria

In the present study, the possible association of thymidine kinase (TK) with mitochondria in Naegleria was investigated by treating growing and differentiating cells with chloramphenicol (CAP), an inhibitor of mitochondrial protein synthesis. In some systems, CAP causes an overproduction of mitochondrial proteins coded for in the nucleus. The present results show that in growing Naegleria , CAP stimulates a dramatic increase in TK activity while growth and division is gradually inhibited. CAP does not stabilize the enzyme in vivo or in vitro. The stimulation is cycloheximide (CHI)‐sensitive and specific since nucleoside phosphotransferase activity does not increase. In cells stimulated to differentiate, CAP does not prevent differentiation or the expected decrease in TK activity. Using polyacrylamide gel electrophoresis, a comparison of TK in mitochondrial and postmitochondrial fractions of CAP‐treated and untreated cells was made. Results suggest some processing of the enzyme, resulting in a slight change in electrophoretic mobility. No mitochondrial TK was found. The stimulation of a cytoplasmic enzyme by CAP suggests a form of mitochondrial control of nuclear transcription for other than mitochondrial proteins. DNA synthesis in CAP‐treated cells was not stimulated, suggesting (since TK and DNA synthesis are usually tightly coupled) an uncoupling of these two events, most likely, at the beginning of the S phase.