Sodium channel activity in leukemia cells is directly controlled by actin polymerization.
The actin cytoskeleton has been shown to be involved in the regulation of sodium-selective channels in non-excitable cells. However, the molecular mechanisms underlying the changes in channel function remain to be defined. In the present work, inside-out patch experiments were employed to elucidate the role of submembranous actin dynamics in the control of sodium channels in human myeloid leukemia K562 cells. We found that the application of cytochalasin D to the cytoplasmic surface of membrane fragments resulted in activation of non-voltage-gated sodium channels of 12 picosiemens conductance. Similar effects could be evoked by addition of the actin-severing protein gelsolin to the bath cytosol-like solution containing 1 microm [Ca(2+)](i). The sodium channel activity induced by disassembly of submembranous microfilaments with cytochalasin D or gelsolin could be abolished by intact actin added to the bath cytosol-like solution in the presence of 1 mm MgCl(2) to induce actin polymerization. In the absence of MgCl(2), addition of intact actin did not abolish the channel activity. Moreover, the sodium currents were unaffected by heat-inactivated actin or by actin whose polymerizability was strongly reduced by cleavage with specific Escherichia coli A2 protease ECP32. Thus, the inhibitory effect of actin on channel activity was observed only under conditions promoting rapid polymerization. Taken together, our data show that sodium channels are directly controlled by dynamic assembly and disassembly of submembranous F-actin.
Negulyaev
Y A
YA
Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Avenue, St. Petersburg 194064, Russia.
Khaitlina
S Y
SY
Hinssen
H
H
Shumilina
E V
EV
Vedernikova
E A
EA
D000199:Actins / Q000378:metabolism*; D015638:Cytochalasin D / Q000494:pharmacology; D018260:Gelsolin / Q000494:pharmacology; D006801:Humans; D020014:K562 Cells; D007938:Leukemia / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D011108:Polymers / Q000378:metabolism; D015222:Sodium Channels / Q000502:physiology*
D000199:Actins; D018260:Gelsolin; D011108:Polymers; D015222:Sodium Channels; D015638:Cytochalasin D; D008274:Magnesium