RRC ID 51324
Author Cho T, Ishii-Kato A, Fukata Y, Nakayama Y, Iida K, Fukata M, Iida H.
Title Coupling of a voltage-gated Ca2+ channel homologue with a plasma membrane H+ -ATPase in yeast.
Journal Genes Cells
Abstract Yeast has a homologue of mammalian voltage-gated Ca2+ channels (VGCCs), enabling the efficient uptake of Ca2+ . It comprises two indispensable subunits, Cch1 and Mid1, equivalent to the mammalian pore-forming α1 and auxiliary α2 /δ subunits, respectively. Unlike the physiological roles of Cch1/Mid1 channels, the regulatory mechanisms of the yeast VGCC homologue remain unclear. Therefore, we screened candidate proteins that interact with Mid1 by an unbiased proteomic approach and identified a plasma membrane H+ -ATPase, Pma1, as a candidate. Mid1 coimmunoprecipitated with Pma1, and Mid1-EGFP colocalized with Pma1-mCherry at the plasma membrane. The physiological relevance of their interaction was determined using the temperature-sensitive mutant, pma1-10. At the nonpermissive temperature, the membrane potential was less negative and Ca2+ uptake was lower in pma1-10 than in wild-type cells. Increased extracellular H+ increased the rate of Ca2+ uptake. Therefore, H+ extrusion by Pma1 may be important for Ca2+ influx through Cch1/Mid1. These results suggest that Pma1 interacts physically with Cch1/Mid1 Ca2+ channels to enhance their activity via its H+ -pumping activity.
Volume 22(1)
Pages 94-104
Published 2017-1-1
DOI 10.1111/gtc.12458
PMID 27935186
MeSH Amino Acid Sequence Calcium / metabolism Calcium Channels / genetics Calcium Channels / metabolism* Cell Membrane / genetics Cell Membrane / metabolism Membrane Glycoproteins / genetics Membrane Glycoproteins / metabolism* Protein Interaction Mapping / methods Proteomics* Proton-Translocating ATPases / genetics Proton-Translocating ATPases / metabolism* Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins / genetics Saccharomyces cerevisiae Proteins / metabolism*
IF 1.922
Times Cited 2
Resource
DNA material pBCT-CCH1H-EGFP (RDB15298) YCpT-MID1-HAx4 (RDB15299) YCpL-MID1-HAx4 (RDB15300) YCpT-MID1-EGFP (RDB15301) YCpT-PMA1-mCherry (RDB15302) YCpT-PMA1 (RDB15303).