Profound differences in potassium current properties of normal and Rous sarcoma virus-transformed chicken embryo fibroblasts.
The membrane currents of chicken embryo fibroblasts (CEFs) transformed by Rous sarcoma virus (RSV) were compared with the currents of their nontransformed counterparts by using the whole-cell patch-clamp technique. In nontransformed CEFs, the main membrane current is a delayed outward K+ current that is sensitive to tetraethylammonium ion but insensitive to 4-aminopyridine. This K+ current is almost independent of the intracellular Ca2+ concentration and becomes completely inactivated at positive membrane potentials with a time constant of about 10 s at +30 mV. In contrast, transformed CEFs exhibit a noninactivating K+ current that strongly depends on the intracellular Ca2+ concentration. This Ca(2+)-dependent K+ current is blocked by the scorpion toxin charybdotoxin with an IC50 value of 19 nM, whereas the K+ current of normal CEFs is insensitive to charybdotoxin (up to 300 nM). The K+ current properties of transformed CEFs were also found after microinjection of purified, enzymatically active pp60v-src into normal CEFs but not after infection of CEFs with the Rous-associated virus RAV5, which lacks the v-src oncogene. Our results suggest that the oncogene product pp60v-src modulates existing K+ channel proteins, leading to profound electrophysiological and pharmacological alterations of the K+ current properties in RSV-transformed CEFs. Furthermore, our experiments identify for the first time K+ channels as possible substrates of pp60v-src.
Repp
H
H
Rudolf-Buchheim-Institut für Pharmakologie, Justus-Liebig-Universität Giessen, Federal Republic of Germany.
Draheim
H
H
Ruland
J
J
Seidel
G
G
Beise
J
J
Presek
P
P
Dreyer
F
F
D000818:Animals; D001030:Apamin / Q000494:pharmacology; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D018999:Charybdotoxin; D002642:Chick Embryo; D004546:Elapid Venoms / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D008845:Microinjections; D015688:Oncogene Protein pp60(v-src) / Q000008:administration & dosage / Q000378:metabolism*; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D012604:Scorpion Venoms / Q000494:pharmacology
D004546:Elapid Venoms; D015221:Potassium Channels; D012604:Scorpion Venoms; D018999:Charybdotoxin; D001030:Apamin; C020273:dendrotoxin; D015688:Oncogene Protein pp60(v-src)