Testing a proposed paradigm shift in analysis of phage DNA packaging

P. Serwer, E. Wright
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引用次数: 8

Abstract

ABSTRACT We argue that a paradigm shift is needed in the analysis of phage DNA packaging. We then test a prediction of the following paradigm shift-engendering hypothesis. The motor of phage DNA packaging has two cycles: (1) the well-known packaging ATPase-driven (type 1) cycle and (2) a proposed back-up, shell expansion/contraction-driven (type 2) cycle that reverses type 1 cycle stalls by expelling accidentally packaged non-DNA molecules. We test the prediction that increasing the cellular concentration of all macromolecules will cause packaging-active capsids to divert to states of hyper-expansion and contraction. We use a directed evolution-derived, 3-site phage T3 mutant, adapted to propagation in concentrated bacterial cytoplasm. We find this prediction correct while discovering novel T3 capsids previously obscure.
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测试在噬菌体DNA包装分析中提出的范式转变
我们认为在噬菌体DNA包装的分析中需要一个范式转变。然后,我们测试了以下范式转换产生假设的预测。噬菌体DNA包装的马达有两个周期:(1)众所周知的包装atp酶驱动(第1型)周期和(2)提议的备用壳膨胀/收缩驱动(第2型)周期,通过排出意外包装的非DNA分子来逆转第1型周期的停滞。我们测试了增加所有大分子的细胞浓度将导致包装活性衣壳转向超膨胀和超收缩状态的预测。我们使用定向进化衍生的3位点噬菌体T3突变体,适应于在浓缩的细菌细胞质中繁殖。我们发现这一预测是正确的,同时发现了以前不为人知的新型T3衣壳。
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