Remote Activation of Host Cell DNA Synthesis in Uninfected Cells Signaled by Infected Cells in Advance of Virus Transmission.
Remote Activation of Host Cell DNA Synthesis in Uninfected Cells Signaled by Infected Cells in Advance of Virus Transmission.
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DOI：
10.1128/jvi.01950-15
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发表时间：
2015-11
影响因子：
5.4
通讯作者：
O'Hare P
中科院分区：
文献类型：
--
作者：
Schmidt N;Hennig T;Serwa RA;Marchetti M;O'Hare P
病毒以不同的方式调节细胞过程和代谢，但这些几乎都是在受感染的细胞本身中进行研究的。在这里，我们研究的空间组织的DNA合成过程中的多轮传输的单纯疱疹病毒（HSV）使用脉冲标记乙炔基核苷酸和叠氮荧光团的环加成。我们报告了一个迄今为止未知的和意想不到的结果病毒-宿主相互作用。与目前对单步生长周期的理解一致，HSV抑制宿主DNA合成并促进细胞内空间隔离区室中的病毒DNA合成。与此形成鲜明对比的是，在单细胞开始的渐进感染过程中，我们观察到感染会诱导远程未感染细胞中细胞DNA复制的明显且明显的刺激。这种诱导的DNA合成在数百个未感染的细胞中观察到，这些细胞位于扩展边界，在进行性感染的周边之外。此外，使用脉冲追踪分析，我们表明，这种激活被维持，导致在感染前宿主DNA合成的传播波不断。当病毒到达并感染这些激活的细胞时，宿主DNA合成被关闭，取而代之的是病毒DNA合成。使用非繁殖病毒或条件培养基，我们证明了一个旁分泌效应未感染的细胞DNA合成在远程细胞不断提前感染。这些发现具有重要的意义，可能具有广泛的适用性，为我们理解病毒感染操纵细胞过程的方式，不仅在受感染的细胞本身，而且现在在远程未感染的细胞，以及宿主DNA合成的机制。重要性我们表明，在由具有进行性细胞-细胞病毒传播的单个颗粒引发的感染期间（即，正常情况下），HSV通过病毒诱导的旁分泌效应物介导，诱导未感染细胞中的宿主DNA合成。该领域没有概念，这一过程发生，工作改变了我们的解释病毒-宿主相互作用在推进感染和理解宿主DNA合成的控制有影响。我们的研究结果证明了化学生物学技术在分析感染过程中的实用性，揭示了在多轮传播与单步生长曲线中检查感染时的不同过程，并揭示了病毒感染中迄今未知的过程，可能与其他病毒（和其他感染因子）以及其他过程（包括转录和蛋白质合成）的远程信号相关。
Viruses modulate cellular processes and metabolism in diverse ways, but these are almost universally studied in the infected cell itself. Here, we study spatial organization of DNA synthesis during multiround transmission of herpes simplex virus (HSV) using pulse-labeling with ethynyl nucleotides and cycloaddition of azide fluorophores. We report a hitherto unknown and unexpected outcome of virus-host interaction. Consistent with the current understanding of the single-step growth cycle, HSV suppresses host DNA synthesis and promotes viral DNA synthesis in spatially segregated compartments within the cell. In striking contrast, during progressive rounds of infection initiated at a single cell, we observe that infection induces a clear and pronounced stimulation of cellular DNA replication in remote uninfected cells. This induced DNA synthesis was observed in hundreds of uninfected cells at the extended border, outside the perimeter of the progressing infection. Moreover, using pulse-chase analysis, we show that this activation is maintained, resulting in a propagating wave of host DNA synthesis continually in advance of infection. As the virus reaches and infects these activated cells, host DNA synthesis is then shut off and replaced with virus DNA synthesis. Using nonpropagating viruses or conditioned medium, we demonstrate a paracrine effector of uninfected cell DNA synthesis in remote cells continually in advance of infection. These findings have significant implications, likely with broad applicability, for our understanding of the ways in which virus infection manipulates cell processes not only in the infected cell itself but also now in remote uninfected cells, as well as of mechanisms governing host DNA synthesis. IMPORTANCE We show that during infection initiated by a single particle with progressive cell-cell virus transmission (i.e., the normal situation), HSV induces host DNA synthesis in uninfected cells, mediated by a virus-induced paracrine effector. The field has had no conception that this process occurs, and the work changes our interpretation of virus-host interaction during advancing infection and has implications for understanding controls of host DNA synthesis. Our findings demonstrate the utility of chemical biology techniques in analysis of infection processes, reveal distinct processes when infection is examined in multiround transmission versus single-step growth curves, and reveal a hitherto-unknown process in virus infection, likely relevant for other viruses (and other infectious agents) and for remote signaling of other processes, including transcription and protein synthesis.