TY - JOUR
T1 - Non-endocytic penetration of core histones into petunia protoplasts and cultured cells
T2 - A novel mechanism for the introduction of macromolecules into plant cells
AU - Rosenbluh, Joseph
AU - Singh, Sunil Kumar
AU - Gafni, Yedidya
AU - Graessmann, Adolf
AU - Loyter, Abraham
PY - 2004/8/30
Y1 - 2004/8/30
N2 - The results of the present work demonstrate that core histones are able to penetrate the plasma membrane of plant cells. Confocal microscopy has revealed that incubation of petunia protoplasts with fluorescently labeled core histones resulted in cell penetration and nuclear import of the externally added histones. Intracellular accumulation was also confirmed by an ELISA-based quantitative method using biotin-labeled histones. Penetration into petunia protoplasts and cultured cells was found to be non-saturable, occurred at room temperature and at 4°C and was not inhibited by Nocodazole. Furthermore, penetration of the biotinylated histone was neither blocked by the addition of an excess of free biotin molecules, nor by non-biotinylated histone molecules. All these results clearly indicate that the observed uptake is due to direct translocation through the cell plasma membrane and does not occur via endocytosis. Our results also show that the histones H2A and H4 were able to mediate penetration of covalently attached BSA molecules demonstrating the potential of the histones as carriers for the delivery of macromolecules into plant cells. To the best of our knowledge, the findings of the present paper demonstrate, for the first time, the activity of cell penetrating proteins (CPPs) in plant cells.
AB - The results of the present work demonstrate that core histones are able to penetrate the plasma membrane of plant cells. Confocal microscopy has revealed that incubation of petunia protoplasts with fluorescently labeled core histones resulted in cell penetration and nuclear import of the externally added histones. Intracellular accumulation was also confirmed by an ELISA-based quantitative method using biotin-labeled histones. Penetration into petunia protoplasts and cultured cells was found to be non-saturable, occurred at room temperature and at 4°C and was not inhibited by Nocodazole. Furthermore, penetration of the biotinylated histone was neither blocked by the addition of an excess of free biotin molecules, nor by non-biotinylated histone molecules. All these results clearly indicate that the observed uptake is due to direct translocation through the cell plasma membrane and does not occur via endocytosis. Our results also show that the histones H2A and H4 were able to mediate penetration of covalently attached BSA molecules demonstrating the potential of the histones as carriers for the delivery of macromolecules into plant cells. To the best of our knowledge, the findings of the present paper demonstrate, for the first time, the activity of cell penetrating proteins (CPPs) in plant cells.
KW - Cell penetrating protein (CPP)
KW - Endocytosis
KW - Histone
KW - Macromolecule delivery
KW - Membrane penetration
KW - Plant protoplast
UR - http://www.scopus.com/inward/record.url?scp=4344609460&partnerID=8YFLogxK
U2 - 10.1016/j.bbamem.2004.06.003
DO - 10.1016/j.bbamem.2004.06.003
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C2 - 15328056
AN - SCOPUS:4344609460
SN - 0005-2736
VL - 1664
SP - 230
EP - 240
JO - Biochimica et Biophysica Acta - Biomembranes
JF - Biochimica et Biophysica Acta - Biomembranes
IS - 2
ER -