Gross K J, Pogo A O
Biochemistry. 1976 May 18;15(10):2082-6. doi: 10.1021/bi00655a008.
The addition of cycloheximide to a thermosensitive conditional yeast mutant (ts-187) before and after transfer to the nonpermissive temperature (36 degrees C) for initiation of protein synthesis produces the uncoupling of the RNA and protein synthetic machineries. Since the drug can produce this relaxation in the presence and absence of protein synthesis, it is concluded that the coupling of protein and RNA synthesis, which a temperature shift produces, is not exclusively related to the inhibition of protein synthesis. Support for this assumption has been obtained using the parental (A364A) strain. Transferring this strain to 36 degrees C produces inhibition of RNA synthesis in the presence of stimulation of protein synthesis. Furthermore, cycloheximide and edeine prevent this inhibtion of RNA synthesis that temperature shift produces. It is, therefore, postulated that this inhibition of RNA synthesis results from the synthesis or activation of a factor(s) elicited by the increase in temperature whose function is to repress the transcriptional apparatus. Cycloheximide or edeine can prevent the function of this repressor-like factor by binding to the factor or by preventing its synthesis. The fact that inhibition of protein synthesis either by cycloheximide action or temperature shift in ts-187 produces inhibition of RNA synthesis in isolated nuclei indicates that, in addition to the aforementioned repressor, other factor(s) having a promoter function may exist. Since a slight inhibition of protein synthesis produces nuclear template restrictions, it is postulated that the promoter-like factor(s) is a polypeptide different from the RNA polymerase and, at least in yeast, has a high turnover.
在将温度敏感型条件性酵母突变体(ts - 187)转移至蛋白质合成起始的非允许温度(36摄氏度)之前和之后,添加放线菌酮会导致RNA和蛋白质合成机制的解偶联。由于该药物在有或无蛋白质合成的情况下均能产生这种解偶联作用,因此可以得出结论,温度变化所导致的蛋白质与RNA合成的偶联并非仅仅与蛋白质合成的抑制相关。使用亲本菌株(A364A)已获得了对这一假设的支持。将该菌株转移至36摄氏度会在蛋白质合成受到刺激的情况下抑制RNA合成。此外,放线菌酮和伊短菌素可阻止温度变化所导致的RNA合成抑制。因此,据推测,这种RNA合成抑制是由温度升高引发的一种或多种因子的合成或激活所致,其功能是抑制转录装置。放线菌酮或伊短菌素可通过与该抑制因子样因子结合或阻止其合成来阻止其功能发挥。在ts - 187中,无论是通过放线菌酮作用还是温度变化抑制蛋白质合成,都会在分离的细胞核中抑制RNA合成,这一事实表明,除了上述抑制因子外,可能还存在其他具有启动子功能的因子。由于对蛋白质合成的轻微抑制会产生核模板限制,因此据推测,这种启动子样因子是一种不同于RNA聚合酶的多肽,并且至少在酵母中具有较高的周转率。