Iborra, F.J., Jackson, D.A. and Cook, P.R.
Notes: To understand if protein translation occurs in the nucleus of eukaryotic cells, the researchers in this study labeled proteins in vivo by bathing permeablized cells in a translation mix containing radiochemical, biotin or fluorescently tagged amino acids. The study found that all three types of label gave similar results. To fluorescently tag proteins, cells were grown, then permeablized, and bathed in a mix of buffer, creatine phosphokinase, phosphocreatine, GTP, tRNA, aminoacyl-tRNA synthetase, MgCl2, protease inhibitors, a 1:10 dilution of Fluorotect Greenlys BODIPY labeled lysine-tRNA complex and 50μM amino acid mixture minus lysine. The study revealed that, contrary to previous beliefs, some proteins are transcribed and translated within the nucleus in a coupled manner. (2744)