Dataset Information

Ligand-dependent activation of Slo family channels is defined by interchangeable cytosolic domains.

ABSTRACT: Large-conductance Ca2+- and voltage-regulated K+ channels (Slo1 BK-type) are controlled by two physiological stimuli, membrane voltage and cytosolic Ca2+. Regulation by voltage is similar to that in voltage-dependent K+ channels, arising from positively charged amino acids primarily within the S4 transmembrane helices. The basis for regulation by Ca2+ remains controversial. One viewpoint suggests that the extensive cytosolic C terminus contains the Ca2+ regulatory machinery, whereas another suggests that the pore-forming module contains the Ca2+-sensing elements. To address this issue, we take advantage of another Slo family member, the pH-regulated homolog Slo3. We reason that if the ligand-sensing apparatus is uniquely associated with a particular domain (either the pore or the cytosolic domain), exchange of those domains between Slo1 and Slo3 should result in exchange of ligand dependence in association with the key domain. The results show that the Slo3 cytosolic module confers pH-dependent regulation on the Slo1 pore module, whereas the Slo1 cytosolic module confers Ca2+-dependent regulation on the Slo3 pore module. Thus, ligand-specific regulation is defined by interchangeable cytosolic regulatory modules.

SUBMITTER: Xia XM

PROVIDER: S-EPMC6729329 | biostudies-literature | 2004 Jun

REPOSITORIES: biostudies-literature

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Ligand-dependent activation of Slo family channels is defined by interchangeable cytosolic domains.

Xia Xiao-Ming XM Zhang Xue X Lingle Christopher J CJ

The Journal of neuroscience : the official journal of the Society for Neuroscience 20040601 24

Large-conductance Ca2+- and voltage-regulated K+ channels (Slo1 BK-type) are controlled by two physiological stimuli, membrane voltage and cytosolic Ca2+. Regulation by voltage is similar to that in voltage-dependent K+ channels, arising from positively charged amino acids primarily within the S4 transmembrane helices. The basis for regulation by Ca2+ remains controversial. One viewpoint suggests that the extensive cytosolic C terminus contains the Ca2+ regulatory machinery, whereas another sugg ...[more]

PMID: 15201331

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Project description:The calcium-gated potassium channel SLO-1 in Caenorhabditis elegans was recently identified as key component for action of emodepside, a new anthelmintic drug with broad spectrum activity. In this study we identified orthologues of slo-1 in Ancylostoma caninum, Cooperia oncophora, and Haemonchus contortus, all important parasitic nematodes in veterinary medicine. Furthermore, functional analyses of these slo-1 orthologues were performed using heterologous expression in C. elegans. We expressed A. caninum and C. oncophora slo-1 in the emodepside-resistant genetic background of the slo-1 loss-of-function mutant NM1968 slo-1(js379). Transformants expressing A. caninum slo-1 from C. elegans slo-1 promoter were highly susceptible (compared to the fully emodepside-resistant slo-1(js379)) and showed no significant difference in their emodepside susceptibility compared to wild-type C. elegans (p = 0.831). Therefore, the SLO-1 channels of A. caninum and C. elegans appear to be completely functionally interchangeable in terms of emodepside sensitivity. Furthermore, we tested the ability of the 5' flanking regions of A. caninum and C. oncophora slo-1 to drive expression of SLO-1 in C. elegans and confirmed functionality of the putative promoters in this heterologous system. For all transgenic lines tested, expression of either native C. elegans slo-1 or the parasite-derived orthologue rescued emodepside sensitivity in slo-1(js379) and the locomotor phenotype of increased reversal frequency confirming the reconstitution of SLO-1 function in the locomotor circuits. A potent mammalian SLO-1 channel inhibitor, penitrem A, showed emodepside antagonising effects in A. caninum and C. elegans. The study combined the investigation of new anthelmintic targets from parasitic nematodes and experimental use of the respective target genes in C. elegans, therefore closing the gap between research approaches using model nematodes and those using target organisms. Considering the still scarcely advanced techniques for genetic engineering of parasitic nematodes, the presented method provides an excellent opportunity for examining the pharmacofunction of anthelmintic targets derived from parasitic nematodes.

Dataset Information

Ligand-dependent activation of Slo family channels is defined by interchangeable cytosolic domains.

Publications

Ligand-dependent activation of Slo family channels is defined by interchangeable cytosolic domains.

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