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Commitment to Lysogeny Is Preceded by a Prolonged Period of Sensitivity to the Late Lytic Regulator Q in Bacteriophage λ

机译:对噬菌体的承诺要先对噬菌体λ中后期的溶酶调节剂Q进行长时间的敏感。

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摘要

A key event in development is the irreversible commitment to a particular cell fate, which may be concurrent with or delayed with respect to the initial cell fate decision. In this work, we use the paradigmatic bacteriophage λ lysis-lysogeny decision circuit to study the timing of commitment. The lysis-lysogeny decision is made based on the expression trajectory of CII. The chosen developmental strategy is manifested by repression of the pR and pL promoters by CI (lysogeny) or by antitermination of late gene expression by Q (lysis). We found that expression of Q in trans from a plasmid at the time of infection resulted in a uniform lytic decision. Furthermore, expression of Q up to 50 min after infection results in lysis of the majority of cells which initially chose lysogenic development. In contrast, expression of Q in cells containing a single chromosomal prophage had no effect on cell growth, indicating commitment to lysogeny. Notably, if the prophage was present in 10 plasmid-borne copies, Q expression resulted in lytic development, suggesting that the cellular phage chromosome number is the critical determinant of the timing of lysogenic commitment. Based on our results, we conclude that (i) the lysogenic decision made by the CI-Cro switch soon after infection can be overruled by ectopic Q expression at least for a time equivalent to one phage life cycle, (ii) the presence of multiple λ chromosomes is a prerequisite for a successful Q-mediated switch from lysogenic to lytic development, and (iii) phage chromosomes within the same cell can reach different decisions.
机译:发展中的关键事件是对特定细胞命运的不可逆转的承诺,这可能与初始细胞命运决定同时发生或延迟发生。在这项工作中,我们使用范式噬菌体λ裂解-溶原性决定电路研究承诺的时机。根据CII的表达轨迹做出裂解溶源性决定。选择的发展策略是通过CI(溶源性)抑制pR和pL启动子或通过Q(裂解)抗终止晚期基因表达来体现的。我们发现感染时质粒中反式Q的表达导致了统一的裂解决定。此外,在感染后长达50分钟的Q表达导致大多数细胞裂解,这些细胞最初选择了溶源性发育。相反,在含有单个染色体噬菌体的细胞中Q的表达对细胞生长没有影响,表明对溶原性的承诺。值得注意的是,如果该噬菌体以10个质粒携带的拷贝形式存在,则Q表达会导致裂解发育,这表明细胞噬菌体的染色体数目是溶源性承诺时间的关键决定因素。根据我们的结果,我们得出结论:(i)感染后不久由CI-Cro开关做出的溶原性决定可以被异位Q表达否决,至少持续一个噬菌体生命周期,(ii)存在多个λ染色体是成功的Q介导的从溶原性向裂解性转化的先决条件,并且(iii)同一细胞内的噬菌体染色体可以达到不同的决定。

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