Nanosecond Electric Pulses Affect a Plant-Specific Kinesin at the Plasma Membrane
Nanosecond Electric Pulses Affect a Plant-Specific Kinesin at the Plasma Membrane
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DOI：
10.1007/s00232-013-9594-z
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发表时间：
2013-12-01
影响因子：
2.4
通讯作者：
Nick, Peter
中科院分区：
文献类型：
--
作者：
Kuehn, Sebastian;Liu, Qiong;Nick, Peter
具有纳秒范围内的高场强和持续时间的电脉冲（nsPEF）对于生物技术和医学应用具有相当大的兴趣。然而，它们的实际细胞作用位点仍在争论中-由于它们的极短的上升时间，nsPEFs被认为主要在细胞内部而不是在质膜上起作用。另一方面，nsPEFs可以诱导膜通透性。我们以植物细胞为模型重新讨论了这个问题。通过在烟草BY-2细胞系中绘制对不同场强和持续时间的nsPEF的细胞响应，我们可以定义不影响短期生存力的处理，使得可以跟踪对处理的生理响应。我们观察到，在这些条件下，一个温和的解体的细胞骨架，受损的膜定位的PIN 1生长素外排转运蛋白和延迟有丝分裂前核定位随后由一个短暂的有丝分裂停滞。为了解决nsPEFs的靶位点，我们利用了植物特异性KCH驱动蛋白，其可以呈现两种不同的状态，具有不同的定位（靠近细胞核或在细胞膜处），驱动不同的细胞功能。我们发现，nsPEFs减少非转化细胞的细胞扩增，但促进过表达KCH的细胞系的扩增。由于细胞伸长和细胞变宽与位于细胞膜上的KCH有关，因此KCH过表达蛋白中的反向反应为nsPEFs也在细胞膜上的直接作用提供了证据。
Electric pulses with high field strength and durations in the nanosecond range (nsPEFs) are of considerable interest for biotechnological and medical applications. However, their actual cellular site of action is still under debate-due to their extremely short rise times, nsPEFs are thought to act mainly in the cell interior rather than at the plasma membrane. On the other hand, nsPEFs can induce membrane permeability. We have revisited this issue using plant cells as a model. By mapping the cellular responses to nsPEFs of different field strength and duration in the tobacco BY-2 cell line, we could define a treatment that does not impinge on short-term viability, such that the physiological responses to the treatment can be followed. We observe, for these conditions, a mild disintegration of the cytoskeleton, impaired membrane localization of the PIN1 auxin-efflux transporter and a delayed premitotic nuclear positioning followed by a transient mitotic arrest. To address the target site of nsPEFs, we made use of the plant-specific KCH kinesin, which can assume two different states with different localization (either near the nucleus or at the cell membrane) driving different cellular functions. We show that nsPEFs reduce cell expansion in nontransformed cells but promote expansion in a line overexpressing KCH. Since cell elongation and cell widening are linked to the KCH localized at the cell membrane, the inverted response in the KCH overexpressor provides evidence for a direct action of nsPEFs, also at the cell membrane.