TY - JOUR
T1 - Receptor-mediated endocytosis and nuclear transport of a transfecting DNA construct
AU - Rosenkranz, Andrey A.
AU - Yachmenev, Sergey V.
AU - Jans, David A.
AU - Serebryakova, Natalya V.
AU - Murav'ev, Vitaly I.
AU - Peters, Reiner
AU - Sobolev, Alexander S.
PY - 1992/1/1
Y1 - 1992/1/1
N2 - A soluble construct consisting of a plasmid carrying the gene of the SV40 large T-antigen and an insulin-poly-l-lysine conjugate is able to selectively transfect PLC/PRF/5 human hepatoma cells which possess insulin receptors. Transfection can be efficiently competed by excess free insulin. To examine intracellular transport of the construct, it was fluorescently labeled and its accumulation on and in cells visualized by video-enhanced microscopy and quantitative confocal laser scanning microscopy. After 2 h at 37 °C, the labeled construct was found predominantly in intracellular acidic compartments, with a substantial portion of fluorescence localized both near and in the cell nucleus. Binding, endocytosis, and nuclear localization of the labeled conjugate could all be competed by excess free insulin, thus indicating that entry of the conjugate into cells was specifically mediated by the insulin receptor.
AB - A soluble construct consisting of a plasmid carrying the gene of the SV40 large T-antigen and an insulin-poly-l-lysine conjugate is able to selectively transfect PLC/PRF/5 human hepatoma cells which possess insulin receptors. Transfection can be efficiently competed by excess free insulin. To examine intracellular transport of the construct, it was fluorescently labeled and its accumulation on and in cells visualized by video-enhanced microscopy and quantitative confocal laser scanning microscopy. After 2 h at 37 °C, the labeled construct was found predominantly in intracellular acidic compartments, with a substantial portion of fluorescence localized both near and in the cell nucleus. Binding, endocytosis, and nuclear localization of the labeled conjugate could all be competed by excess free insulin, thus indicating that entry of the conjugate into cells was specifically mediated by the insulin receptor.
UR - http://www.scopus.com/inward/record.url?scp=0026719220&partnerID=8YFLogxK
U2 - 10.1016/0014-4827(92)90441-A
DO - 10.1016/0014-4827(92)90441-A
M3 - Article
C2 - 1312009
AN - SCOPUS:0026719220
SN - 0014-4827
VL - 199
SP - 323
EP - 329
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -