Leipold B
J Virol. 1977 Feb;21(2):445-50. doi: 10.1128/JVI.21.2.445-450.1977.
The polyamine spermidine has recently been reported to be a substantial component of the RNA phage particle. Its effect on the isolated RNA-A protein complex of the phage MS2 is investigated here. This complex infects intact Escherichia coli cells via F-pili, as does the whole phage. It is shown that the infectivity of the complex on intact E. coli cells was enhanced by incubation with spermidine. Optimal stimulation (20-fold) of the complex infectivity was achieved by incubation with 3 x 10(-4) M spermidine for 20 to 30 min at 37 degrees C. This gave a more compact structure to the complex, as could be seen by its faster sedimentation in sucrose gradients. Although spermidine and Mg2+ are known to partially replace one another in several systems, no enhancement of the infectivity of the complex, but only its considerably faster sedimentation in sucrose gradients, occurred after incubation with 3 x 10(-4) M Mg2+. Only if the Mg2+ concentration was raised by more than one order of magnitude could increased infectivity of the complex be observed. At concentrations of spermidine and Mg2+ that maximally stimulated the infectivity of the complex on intact E. coli cells, no increase in infectivity of phenol-extracted RNA to E. coli spheroplasts was detected. From these in vitro results, the role of the polyamine spermidine in the RNA phage particle for the infecting, RNA-A protein complex molecules in phage infection is discussed.
最近有报道称,多胺亚精胺是RNA噬菌体颗粒的重要组成部分。本文研究了其对噬菌体MS2分离的RNA-A蛋白复合物的影响。该复合物通过F菌毛感染完整的大肠杆菌细胞,整个噬菌体也是如此。结果表明,与亚精胺孵育可增强复合物对完整大肠杆菌细胞的感染性。在37℃下用3×10⁻⁴M亚精胺孵育20至30分钟,可实现复合物感染性的最佳刺激(20倍)。这使复合物的结构更紧凑,从其在蔗糖梯度中更快的沉降速度可见一斑。尽管已知在几个系统中亚精胺和Mg²⁺可部分相互替代,但用3×10⁻⁴M Mg²⁺孵育后,复合物的感染性并未增强,只是其在蔗糖梯度中的沉降速度明显加快。只有当Mg²⁺浓度提高一个数量级以上时,才能观察到复合物感染性的增加。在能最大程度刺激复合物对完整大肠杆菌细胞感染性的亚精胺和Mg²⁺浓度下,未检测到酚提取的RNA对大肠杆菌原生质体感染性的增加。基于这些体外实验结果,讨论了多胺亚精胺在RNA噬菌体颗粒中对噬菌体感染过程中感染性RNA-A蛋白复合物分子的作用。