Yeast K1 killer toxin forms ion channels in sensitive yeast spheroplasts and in artificial liposomes.
The patch-clamp technique was used to examine the plasma membranes of sensitive yeast spheroplasts exposed to partially purified killer toxin preparations. Asolectin liposomes in which the toxin was incorporated were also examined. Excised inside-out patches from these preparations often revealed at 118 pS conductance appearing in pairs. The current through this conductance flickered rapidly among three states: dwelling mostly at the unit-open state, less frequently at the two-unit-open state, and more rarely at the closed state. Membrane voltages from -80 to 80 mV had little influence on the opening probability. The current reversed near the equilibrium potential of K+ in asymmetric KCl solutions and also reversed near O mV at symmetric NaCl vs. KCl solutions. The two levels of the conductance were likely due to the toxin protein, as treatment of spheroplasts or liposomes with extracellular protein preparations from isogenic yeasts deleted for the toxin gene gave no such conductance levels. These results show that in vivo the killer-toxin fraction can form a cation channel that seldom closes regardless of membrane voltage. We suggest that this channel causes the death of sensitive yeast cells.
Laboratory of Molecular Biology, University of Wisconsin-Madison 53706.
B
B
Martinac
H
H
Zhu
A
A
Kubalski
X L
XL
Zhou
M
M
Culbertson
H
H
Bussey
C
C
Kung
D002462:Cell Membrane / Q000502:physiology; D004553:Electric Conductivity; D005656:Fungal Proteins / Q000378:metabolism*; D007473:Ion Channels / Q000502:physiology*; D055646:Killer Factors, Yeast; D008081:Liposomes; D008564:Membrane Potentials; D009183:Mycotoxins / Q000378:metabolism* / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins; D013104:Spheroplasts / Q000502:physiology
D005656:Fungal Proteins; D007473:Ion Channels; C527157:KHR1 protein, S cerevisiae; D055646:Killer Factors, Yeast; D008081:Liposomes; D009183:Mycotoxins; D029701:Saccharomyces cerevisiae Proteins