Induction of multiple flagella in Naegleria: Requirements for RNA and protein synthesis

摘要

AbstractThe induction of multiple flagella by a heat shock was used to examine the role of RNA and protein synthesis in the regulation of the number of flagella produced during the differentiation of the amebo‐flagellate Naegleria gruberi. Control cells differentiating at 25.0°C produce an average of 2.1 flagella per flagellate (f/F), whereas cells exposed to 38.2°C from 35 minutes until 72 minutes after initiation produce an average of approximately 5 f/F. Heat shock was found to prevent completion of the RNA synthesis essential for flagellum formation, even though both RNA and protein synthesis continued at 38.2°C. A delay of two minutes for every one minute of heat shock was seen in both the formation of flagella (T50) and the completion of essential RNA synthesis for heat shocks of one to ten minutes applied beginning 35 minutes after initiation. Times at 38.2°C of ten minutes to 45 minutes produced a delay of approximately 0.6 minutes for each minute of heat shock, whereas shocks longer than 47 minutes prevented flagellum formation. The times from completion of RNA synthesis until completion of protein synthesis or flagella formation were found to be approximately 15 minutes and approximately 32 minutes, respectively. This was true in control cells as well as in cells heat shocked for up to 45 minutes. The fact that heat shock delayed completion of RNA synthesis without affecting the time for completion of protein synthesis or flagella formation suggests that heat shock acts at some step related to the completion of transcription. Short heat shocks, 30 seconds to five minutes, were observed to prevent flagellum formation in cells that had completed RNA synthesis if additional RNA synthesis was inhibited. In contrast, short heat shocks had little effect on cells that had completed protein synthesis, even if additional protein synthesis was inhibited. Two alternative hypotheses for the mechanism of heat shock delay of transcription are discussed. One hypothesis involves a direct effect of high temperature on transcription, and the other postulates a temperature‐sensitive protein necessary for flagellum fo