caa a22 4500 606209115 CHVBK 20210128101026.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s00425-014-2216-x doi (NATIONALLICENCE)springer-10.1007/s00425-014-2216-x Do phosphoinositides regulate membrane water permeability of tobacco protoplasts by enhancing the aquaporin pathway? [Elektronische Daten] [Xiaohong Ma, Arava Shatil-Cohen, Shifra Ben-Dor, Noa Wigoda, Imara Perera, Yang Im, Sofia Diminshtein, Ling Yu, Wendy Boss, Menachem Moshelion, Nava Moran] 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 12-30μms−1, while in the controls only by 3-4μms−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. Springer-Verlag Berlin Heidelberg, 2014 BY2 nationallicence Lipid nationallicence Osmotic water permeability coefficient nationallicence P f nationallicence Signaling nationallicence Synthetic-biology nationallicence AtPIPs, NtPIPs, SlPIPs, SsAQPs : Aquaporins of the PIP family from Arabidopsis, tobacco, tomato, Samanea, respectively nationallicence BY2 (NT1) : Bright Yellow-2, tobacco cultured cells nationallicence DAG : Diacylglycerol nationallicence DGK : DAG kinase nationallicence HsPIPKIα : Human PI(4)P 5-kinase type Iα nationallicence InsP3 : Inositol (1,4,5)trisphosphate nationallicence KAT1/SKOR : Arabidopsis K+-influx/K+-release channel nationallicence LKT1 : Tomato K+-influx channel nationallicence MIPs (PIP1s, PIP2s), TIPs, NIPs, SIPs, XIPs : Major Intrinsic membrane Proteins (Plant aquaporins families) nationallicence HsPtase : Human InsP3 5-phosphatase nationallicence PA : Propionic acid nationallicence PC : Phosphatidylcholine nationallicence P f : Osmotic permeability nationallicence P fFit : Home-made program for P f determination through hypotonic assays and data fitting nationallicence PI(s) : Phosphoinositide(s) nationallicence PIP : Phosphatidylinositol phosphate, in the context of membrane phospholipids nationallicence PIP(s) : Aquaporin(s), in the context of a gene or protein family nationallicence PIPK : HsPIPKIα nationallicence PIPK cells : PIPK-transformed tobacco cells nationallicence PLC : Phospholipase C nationallicence PLD : Phospholipase D nationallicence Ptase cells : HsPtase-transformed tobacco cells nationallicence PtdIns : Phosphatidylinositol nationallicence PtdInsP2 : Phosphatidylinositol bisphosphate phosphorylated at positions (4, 5), unless otherwise indicated nationallicence Ma Xiaohong The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Shatil-Cohen Arava The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Ben-Dor Shifra Bioinformatics and Biological Computing Unit, Weizmann Institute of Science, 76100, Rehovot, Israel aut Wigoda Noa The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Perera Imara Department of Plant and Microbial Biology, North Carolina State University, 27695-7649, Raleigh, NC, USA aut Im Yang Department of Plant and Microbial Biology, North Carolina State University, 27695-7649, Raleigh, NC, USA aut Diminshtein Sofia The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Yu Ling The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Boss Wendy Department of Plant and Microbial Biology, North Carolina State University, 27695-7649, Raleigh, NC, USA aut Moshelion Menachem The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Moran Nava The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut Planta Springer Berlin Heidelberg 241/3(2015-03-01), 741-755 0032-0935 241:3<741 2015 241 425 https://doi.org/10.1007/s00425-014-2216-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00425-014-2216-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ma Xiaohong The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Shatil-Cohen Arava The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Ben-Dor Shifra Bioinformatics and Biological Computing Unit, Weizmann Institute of Science, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Wigoda Noa The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Perera Imara Department of Plant and Microbial Biology, North Carolina State University, 27695-7649, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Im Yang Department of Plant and Microbial Biology, North Carolina State University, 27695-7649, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Diminshtein Sofia The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Yu Ling The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Boss Wendy Department of Plant and Microbial Biology, North Carolina State University, 27695-7649, Raleigh, NC, USA aut NATIONALLICENCE 700 1- Moshelion Menachem The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 700 1- Moran Nava The Robert H. Smith Faculty of Agriculture Food and Environment, The Robert H. Smith Institute for Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, 76100, Rehovot, Israel aut NATIONALLICENCE 773 0- Planta Springer Berlin Heidelberg 241/3(2015-03-01), 741-755 0032-0935 241:3<741 2015 241 425