Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Oct 25;25(21):11450.
doi: 10.3390/ijms252111450.

High-Affinity Plasma Membrane Ca2+ Channel Cch1 Modulates Adaptation to Sodium Dodecyl Sulfate-Triggered Rise in Cytosolic Ca2+ Concentration in Ogataea parapolymorpha

Maria Kulakova  1 , Maria Pakhomova  1 , Victoria Bidiuk  1 , Alexey Ershov  1 , Alexander Alexandrov  1 , Michael Agaphonov  1
Affiliations

High-Affinity Plasma Membrane Ca2+ Channel Cch1 Modulates Adaptation to Sodium Dodecyl Sulfate-Triggered Rise in Cytosolic Ca2+ Concentration in Ogataea parapolymorpha

Maria Kulakova et al. Int J Mol Sci. .

Abstract

The cytosolic calcium concentration ([Ca2+]cyt) in yeast cells is maintained at a low level via the action of different transporters sequestrating these cations in the vacuole. Among them, the vacuolar Ca2+ ATPase Pmc1 crucially contributes to this process. Its inactivation in Ogataea yeasts was shown to cause sodium dodecyl sulfate (SDS) hypersensitivity that can be alleviated by the inactivation of the plasma membrane high-affinity Ca2+ channel Cch1. Here, we show that SDS at low concentrations induces a rapid influx of external Ca2+ into cells, while the plasma membrane remains impermeable for propidium iodide. The inactivation of Pmc1 disturbs efficient adaptation to this activity of SDS. The inactivation of Cch1 partially restores the ability of pmc1 mutant cells to cope with an increased [Ca2+]cyt that correlates with the suppression of SDS hypersensitivity. At the same time, Cch1 is unlikely to be directly involved in SDS-induced Ca2+ influx, since its inactivation does not decrease the amplitude of the rapid [Ca2+]cyt elevation in the pmc1-Δ mutant. The obtained data suggest that the effects of CCH1 inactivation on SDS sensitivity and coping with increased [Ca2+]cyt are related to an additional Cch1 function beyond its direct involvement in Ca2+ transport.

Keywords: Hog1 phosphorylation; Ogataea; genetically encoded Ca2+ indicator; high-affinity Ca2+ uptake system; vacuolar Ca2+ ATPase; voltage-gated calcium channel; yeast.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Change in CFU numbers (survival rate) after 10′ and 60′ incubation in presence of 0.01% SDS. Log., logarithmic cultures; Stat., stationary cultures; WT, DL5-LC strain; cch1-Δ, DL5-cch1 strain; pmc1-Δ, DL5-pmc1-LC strain; pmc1-Δ cch1-Δ, DL5-pmc1-cch1 strain. For details, see Section 4. *—p-value < 0.005.
Figure 2
Figure 2
Distribution of fluorescence (FL) of DL5-LC (WT) and DL5-pmc1-LC (pmc1-Δ) cells after PI staining. Red line “SDS”, cells after 1 h incubation with 0.01% SDS; blue line “C”, untreated control cells; black line “B”, cells inactivated by boiling. Comparison of three replicates is represented in Supplementary Figure S1.
Figure 3
Figure 3
Dynamics of median FL450/FL525 values of cells of DL5-LC (WT), DL5-cch1 (cch1-Δ), DL5-pmc1-LC (pmc1-Δ) and DL5-pmc1-cch1 strains expressing GEM-GECO. (A), response of DL5-LC cells in logarithmic culture to different SDS concentrations; (B), response of cells in logarithmic cultures to 100 mM CaCl2; (C), response of cells in logarithmic cultures to 0.01% SDS; (D), response of cells in stationary cultures to 0.01% SDS; (E), response of cells in logarithmic cultures to 0.004% SDS; (F), response of cells in stationary cultures to 0.004% SDS. Median FL450/FL525 values of 104 cells obtained from 3 replicates were averaged, and standard deviations were calculated.
Figure 4
Figure 4
Distributions of FL450/FL525 in exponentially grown cultures of DL5-LC (WT) and DL5-pmc1-LC (pmc1-Δ) strains before (-) or after 10′ incubation with 0.01% SDS in absence (SDS) or presence of 10 mM EDTA (SDS + EDTA) or 20 mM EGTA (SDS + EGTA). Median values are indicated by bars.
Figure 5
Figure 5
SDS-PAGE and immunoblotting of phosphorylated (p38PP) and total Hog1 (Hog1). Exponential cultures were supplemented with 0.004% SDS (4), 0.008% SDS (8) or 0.6 M NaCl and incubated for 3, 10 or 20 min. Untreated cells (-) were used as reference. hog1-Δ mutant was used as negative control. Panel “PMC1”—DL5-LC strain; panel “pmc1-Δ”—DL5-pmc1-LC strain; panel “pmc1-Δ vs. PMC1”—comparison of DL5-pmc1-LC and DL5-LC strains after 20 min incubation with SDS.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Lange M., Peiter E. Calcium Transport Proteins in Fungi: The Phylogenetic Diversity of Their Relevance for Growth, Virulence, and Stress Resistance. Front. Microbiol. 2020;10:3100. doi: 10.3389/fmicb.2019.03100. - DOI - PMC - PubMed
    1. Dong X.-Y. Calcium Ion Channels in Saccharomyces Cerevisiae. J. Fungi. 2023;9:524. doi: 10.3390/jof9050524. - DOI - PMC - PubMed
    1. Paidhungat M., Garrett S. A Homolog of Mammalian, Voltage-Gated Calcium Channels Mediates Yeast Pheromone-Stimulated Ca2+ Uptake and Exacerbates the Cdc1(Ts) Growth Defect. Mol. Cell. Biol. 1997;17:6339–6347. doi: 10.1128/MCB.17.11.6339. - DOI - PMC - PubMed
    1. Fischer M., Schnell N., Chattaway J., Davies P., Dixon G., Sanders D. The Saccharomyces Cerevisiae CCH1 Gene Is Involved in Calcium Influx and Mating. FEBS Lett. 1997;419:259–262. doi: 10.1016/S0014-5793(97)01466-X. - DOI - PubMed
    1. van der Aart Q.J., Kleine K., Steensma H.Y. Sequence Analysis of the 43 Kb CRM1-YLM9-PET54-DIE2-SMI1-PHO81-YHB4-PFK1 Region from the Right Arm of Saccharomyces Cerevisiae Chromosome VII. Yeast. 1996;12:385–390. doi: 10.1002/(SICI)1097-0061(19960330)12:4<385::AID-YEA910>3.0.CO;2-G. - DOI - PubMed

MeSH terms

LinkOut - more resources