Nanosecond electric pulses affect a plant-specific kinesin at the plasma membrane.
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.
Botanical Institute, Karlsruhe Institute of Technology, Kaiserstr. 2, 76128, Karlsruhe, Germany, sebastian.kuehn@kit.edu.
Sebastian
S
Kühn
Qiong
Q
Liu
Christian
C
Eing
Wolfgang
W
Frey
Peter
P
Nick
D027682:Cation Transport Proteins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D004558:Electric Stimulation; D015870:Gene Expression; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis; D018521:Plant Physiological Phenomena; D010940:Plant Proteins / Q000378:metabolism*
D027682:Cation Transport Proteins; D010940:Plant Proteins; D016547:Kinesins