Apirion D, Neil J, Ko T S, Watson N
Genetics. 1978 Sep;90(1):19-35. doi: 10.1093/genetics/90.1.19.
To determine if proteins RNase III and rho, both of which can determine the 3' ends of RNA molecules, can complement each other, double mutants defective in these two factors were constructed. In all cases (four rho mutations tested) the double mutants were viable at lower temperatures, but were unable to grow at higher temperatures at which both of the parental strains grew. Genetic analyses suggested that the combinations of the rnc rho (RNase III-Rho-) mutations was necessary and probably sufficient to confer temperature sensitivity on carrier strains. Physiological studies showed that synthesis and maturation of rRNA, which is greatly affected by RNase III, as well as other RNAs, was indistinguishable in rnc rho strains as compared to rnc rho+ strains, thus suggesting that RNase III and rho do not complement one another in determining the 3' ends of RNA molecules. In rnc rho strains, however, the newly synthesized rRNA failed to accumulate. Thus, decay of rRNA could be the reason for the temperature sensitivity of the double mutant strains. These experiments suggest that RNase III and rho can both protect rRNA from degradation by cellular ribonucleases. They also point to the possibility that the nucleotide sequences involved in the determination of the 3' ends of RNA molecules by these two factors are not identical.
为了确定两种都能决定RNA分子3'末端的蛋白质——核糖核酸酶III(RNase III)和rho因子是否能相互补充,构建了这两种因子存在缺陷的双突变体。在所有情况下(测试了四种rho突变),双突变体在较低温度下能够存活,但在两种亲代菌株都能生长的较高温度下无法生长。遗传分析表明,rnc rho(核糖核酸酶III - Rho -)突变的组合对于赋予载体菌株温度敏感性是必要的,并且可能是充分的。生理学研究表明,与rnc rho +菌株相比,在rnc rho菌株中,受核糖核酸酶III极大影响的rRNA以及其他RNA的合成和成熟没有区别,这表明核糖核酸酶III和rho因子在决定RNA分子的3'末端时不能相互补充。然而,在rnc rho菌株中,新合成的rRNA无法积累。因此,rRNA的降解可能是双突变体菌株温度敏感性的原因。这些实验表明,核糖核酸酶III和rho因子都可以保护rRNA不被细胞核糖核酸酶降解。它们还指出了这样一种可能性,即这两种因子在决定RNA分子3'末端时所涉及的核苷酸序列并不相同。