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Journal article

Dynamic calcium-mediated stress response and recovery signatures in the fungal pathogen, Candida albicans

Abstract:

Calcium (Ca2+) is an important second messenger for activating stress response signaling and cell adaptation in eukaryotic cells yet intracellular Ca2+-dynamics in fungi are poorly understood due to lack of effective real-time Ca2+ reporters. We engineered the GCaMP6f construct for use in the fungal pathogen, Candida albicans, and used live-cell imaging to observe both dynamic Ca2+ spiking and slower changes in non-spiking Ca2+-GCaMP signals elicited by stress or gene deletion. Short-term exposure to membrane, osmotic or oxidative stress generated immediate stress-specific responses and repeated exposure revealed differential recovery signatures. Osmotic stress caused yeast cell shrinkage and no adaptation response, where Ca2+-GCaMP spiking was inhibited by 1 M NaCl but not by 0.666 M CaCl2. Treatment with sodium dodecylsulfate (SDS) caused a spike-burst, raised the non-spiking Ca2+-GCaMP signals, and caused significant cell death, but surviving cells adapted over subsequent exposures. Treatment with 5 mM H2O2 abolished spiking and caused transient non-GCaMP-related autofluorescence, but cells adapted such that spiking returned and autofluorescence diminished on repeated exposure. Adaptation to H2O2 was dependent on Cap1, extracellular Ca2+, and calcineurin but not on its downstream target, Crz1. Ca2+-dynamics were not affected by H2O2 in the hog1Δ or yvc1Δ mutants, suggesting a pre-adapted, resistant state, possibly due to changes in membrane permeability. Live-cell imaging of Ca2+-GCaMP responses in individual cells has, therefore, revealed the dynamics of Ca2+-influx, signaling and homeostasis, and their role in the temporal stress response signatures of C. albicans.

Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1128/mbio.01157-23

Authors


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Role:
Author
ORCID:
0000-0001-7127-2642
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Role:
Author
ORCID:
0000-0002-2765-0594
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Role:
Author
ORCID:
0000-0001-8902-1892
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Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Sub department:
Plant Sciences
Oxford college:
Pembroke College
Role:
Author
ORCID:
0000-0002-8942-6897
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Role:
Author
ORCID:
0000-0003-2038-3611


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Funder identifier:
https://ror.org/029chgv08
Grant:
WT204909/Z/16/Z


Publisher:
American Society for Microbiology
Journal:
mBio More from this journal
Volume:
14
Issue:
5
Article number:
e0115723
Publication date:
2023-09-26
Acceptance date:
2023-08-07
DOI:
EISSN:
2150-7511
ISSN:
2161-2129
Pmid:
37750683


Language:
English
Keywords:
Pubs id:
1536814
Local pid:
pubs:1536814
Deposit date:
2024-09-05

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