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Do phosphoinositides regulate membrane water permeability of tobacco protoplasts by enhancing the aquaporin pathway?


Main conclusion

Enhancing the membrane content of PtdInsP 2 , the already-recognized protein-regulating lipid, increased the osmotic water permeability of tobacco protoplasts, apparently by increasing the abundance of active aquaporins in their membranes.

While phosphoinositides are implicated in cell volume changes and are known to regulate some ion channels, their modulation of aquaporins activity has not yet been reported for any organism. To examine this, we compared the osmotic water permeability (P f) of protoplasts isolated from tobacco (Nicotiana tabacum) cultured cells (NT1) with different (genetically lowered or elevated relative to controls) levels of inositol trisphosphate (InsP3) and phosphatidyl inositol [4,5] bisphosphate (PtdInsP2). To achieve this, the cells were transformed with, respectively, the human InsP3 5-phosphatase (‘Ptase cells’) or human phosphatidylinositol (4) phosphate 5-kinase (‘PIPK cells’). The mean P f of the PIPK cells was several-fold higher relative to that of controls and Ptase cells. Three results favor aquaporins over the membrane matrix as underlying this excessive P f: (1) transient expression of the maize aquaporin ZmPIP2;4 in the PIPK cells increased P f by 12–30 μm s−1, while in the controls only by 3–4 μm s−1. (2) Cytosol acidification—known to inhibit aquaporins—lowered the P f in the PIPK cells down to control levels. (3) The transcript of at least one aquaporin was elevated in the PIPK cells. Together, the three results demonstrate the differences between the PIPK cells and their controls, and suggest a hitherto unobserved regulation of aquaporins by phosphoinositides, which could occur through direct interaction or indirect phosphoinositides-dependent cellular effects.

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Fig. 1
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Fig. 5



Aquaporins of the PIP family from Arabidopsis, tobacco, tomato, Samanea, respectively

BY2 (NT1):

Bright Yellow-2, tobacco cultured cells




DAG kinase


Human PI(4)P 5-kinase type Iα

InsP3 :

Inositol (1,4,5)trisphosphate


Arabidopsis K+-influx/K+-release channel


Tomato K+-influx channel

MIPs (PIP1s, PIP2s), TIPs, NIPs, SIPs, XIPs:

Major Intrinsic membrane Proteins (Plant aquaporins families)


Human InsP3 5-phosphatase


Propionic acid



P f :

Osmotic permeability

P fFit:

Home-made program for P f determination through hypotonic assays and data fitting




Phosphatidylinositol phosphate, in the context of membrane phospholipids


Aquaporin(s), in the context of a gene or protein family



PIPK cells:

PIPK-transformed tobacco cells


Phospholipase C


Phospholipase D

Ptase cells:

HsPtase-transformed tobacco cells



PtdInsP2 :

Phosphatidylinositol bisphosphate phosphorylated at positions (4, 5), unless otherwise indicated


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We thank Prof. M. Maeshima of Nagoya University (Japan) and Prof. R. Kaldenhoff of Darmstadt University of Technology (Germany) for their generous gifts of antibodies, Ms. Lea Naveh for technical help and Ms. Maital Ashkenazi for help with bioinformatics at the MS initial phase. This work was supported by The US-Israel Binational Science Foundation (Grant No. 2000191 to NM and WFB), The Israel Science Foundation (Grant Nos. 550/01, to NM, 953/07 to MM and NM), the US National Science Foundation (to IYP # MCB0718452) and the North Carolina Agricultural Research Service (to WFB).

Conflict of interest

The authors declare that they have no conflict of interest.

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Correspondence to Nava Moran.

Additional information

X. Ma and A. Shatil-Cohen contributed equally to the work.

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Ma, X., Shatil-Cohen, A., Ben-Dor, S. et al. Do phosphoinositides regulate membrane water permeability of tobacco protoplasts by enhancing the aquaporin pathway?. Planta 241, 741–755 (2015).

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  • BY2
  • Lipid
  • Osmotic water permeability coefficient
  • P f
  • Signaling
  • Synthetic-biology