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Journal of Bacteriology, January 1999, p. 291-297, Vol. 181, No. 1
Plant Laboratory,
Received 24 August 1998/Accepted 19 October 1998
We have used the patch clamp technique to characterize whole-cell
currents in spheroplasts isolated from a trk1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Divalent Cation Block of Inward Currents and
Low-Affinity K+ Uptake in Saccharomyces
cerevisiae
trk2
strain of Saccharomyces cerevisiae which lacks high- and
moderate-affinity K+ uptake capacity. In solutions in which
extracellular divalent cation concentrations were 0.1 mM, cells
exhibited a large inward current. This current was not the result of
increasing leak between the glass pipette and membrane, as there was no
effect on the outward current. The inward current comprised both
instantaneous and time-dependent components. The magnitude of the
inward current increased with increasing extracellular K+
and negative membrane potential but was insensitive to extracellular anions. Replacing extracellular K+ with Rb+,
Cs+, or Na+ only slightly modulated the
magnitude of the inward current, whereas replacement with
Li+ reduced the inward current by approximately 50%, and
tetraethylammonium (TEA+) and choline were relatively
impermeant. The inward current was blocked by extracellular
Ca2+ and Mg2+ with apparent
Kis (at
140 mV) of 363 ± 78 and 96 ± 14 µM, respectively. Furthermore, decreasing cytosolic
K+ increased the magnitude of the inward current
independently of the electrochemical driving force for K+
influx, consistent with regulation of the inward current by cytosolic K+. Uptake of 86Rb+ by intact
trk1
trk2
cells was inhibited by extracellular
Ca2+ with a Ki within the range
observed for the inward current. Furthermore, increasing extracellular
Ca2+ from 0.1 to 20 mM significantly inhibited the growth
of these cells. These results are consistent with those of the patch
clamp experiments in suggesting that low-affinity uptake of alkali
cations in yeast is mediated by a transport system sensitive to
divalent cations.
*
Corresponding author. Mailing address: Department of
Biology, University of York, P.O. Box 373, York YO1 5YW, United
Kingdom. Phone: 01904 432854. Fax: 01904 434317. E-mail:
skr4{at}york.ac.uk.
Journal of Bacteriology, January 1999, p. 291-297, Vol. 181, No. 1
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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