Translocation of NLS–BSA conjugates into nuclei of permeabilized mammalian cells can be supported by protoplast extract

FEBS Letters

Friedler, Assaf¹  Loyter, Abraham²  Stanhill, Ariel²  Zakai, Nehama²  Gilon, Chaim¹  Broder, Yehoshua C² 

[1] Department of Organic Chemistry, The Hebrew University of Jerusalem, 91904 Jerusalem, Israel;Department of Biological Chemistry, The Institute of Life Sciences, The Hebrew University of Jerusalem, 91904 Jerusalem, Israel

关键词: Nuclear import;    Petunia extract;    Permeabilized cell;    Plant protoplast;   

DOI  :  10.1016/S0014-5793(97)00855-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An heterologous experimental system, which allows the study of the yet unknown cytosolic factors involved in nuclear import of nuclear localization signal (NLS)-containing proteins in plants, has been established. The ability of plant cell extract to substitute mammalian cytosol and to promote translocation of NLS-containing proteins into nuclei of permeabilized HeLa cells was demonstrated. The results described in the present work show that nuclear import of fluorescently labeled BSA conjugates bearing the NLS sequence of SV40 large T antigen could be supported by petunia cell cytoplasmic extract. This heterologous system shows the characteristic features of the homologous mammalian system, namely, is ATP dependent and is inhibited by WGA, GTPγS as well as by non-fluorescent NLS–BSA conjugates. The system described here offers an experimental method to study and characterise cytosolic factors which are required for nuclear import in plants.

【 授权许可】

Unknown