Intercellular communication is reduced by TPA and Ki-ras p21 in quiescent, but not proliferating, NRK cells.
We investigated whether the growth state of NRK cells (proliferating or quiescent by serum deprivation) affected the ability of oncogenic Ki-ras p21 and the protein kinase C activator, 12-O-tetradecanoylphorbol-13-acetate (TPA), to alter gap junctional communication. We evaluated gap junctional permeance by rate analysis of the transfer of a fluorescent dye, Lucifer Yellow, between cell pairs. We found that while the gap junctions of proliferating NRK cells were unresponsive to both TPA and to Ki-ras p21, junctional communication in quiescent cells was significantly inhibited by brief exposures to 100 ng/ml TPA. Furthermore, activity of Ki-ras p21 2 h prior to TPA exposure enhanced the inhibitory effect of TPA in quiescent cells. Junctional sensitivity to TPA was transient, with inhibition of junctional communication detected at 10 min and refractory after 60 min of continuous exposure. The suppression of junctional communication by TPA was completely prevented if the oncogenic p21 had been active for a longer period of time (48 h). The application of a phorbol ester derivative (4 alpha-PDD), which does not activate protein kinase C, did not affect the ability of quiescent cells to communicate. From these results we conclude that there is a cell-state dependence of junctional sensitivity to TPA in NRK cells and that ras p21 activity potentiates the junctional response to TPA. One interesting possibility is that this involved a cell-cycle effect.
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108.
A F
AF
Paulson
R G
RG
Johnson
M M
MM
Atkinson
D000818:Animals; D002273:Carcinogens / Q000494:pharmacology; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division; D002460:Cell Line; D002461:Cell Line, Transformed; D011905:Genes, ras; D007365:Intercellular Junctions / Q000187:drug effects / Q000502:physiology*; D007668:Kidney; D007700:Kinetics; D007708:Kirsten murine sarcoma virus / Q000235:genetics; D015689:Oncogene Protein p21(ras) / Q000378:metabolism*; D010703:Phorbol Esters / Q000494:pharmacology; D051381:Rats; D013696:Temperature; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D002273:Carcinogens; D010703:Phorbol Esters; C008876:phorbol-12,13-didecanoate; D015689:Oncogene Protein p21(ras); D013755:Tetradecanoylphorbol Acetate