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PMID: 27021331 Published · epublish English Journal Article

Highly Dynamic and Specific Phosphatidylinositol 4,5-Bisphosphate, Septin, and Cell Wall Integrity Pathway Responses Correlate with Caspofungin Activity against Candida albicans.

Antimicrobial agents and chemotherapy ·Vol. 60 ·No. 6 ·2016-00-00 ·Pages 3591-600

Badrane H, Nguyen MH, Clancy CJ

Abstract

Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] activates the yeast cell wall integrity pathway. Candida albicans exposure to caspofungin results in the rapid redistribution of PI(4,5)P2 and septins to plasma membrane foci and subsequent fungicidal effects. We studied C. albicans PI(4,5)P2 and septin dynamics and protein kinase C (PKC)-Mkc1 cell wall integrity pathway activation following exposure to caspofungin and other drugs. PI(4,5)P2 and septins were visualized by live imaging of C. albicans cells coexpressing green fluorescent protein (GFP)-pleckstrin homology (PH) domain and red fluorescent protein-Cdc10p, respectively. PI(4,5)P2 was also visualized in GFP-PH domain-expressing C. albicans mkc1 mutants. Mkc1p phosphorylation was measured as a marker of PKC-Mkc1 pathway activation. Fungicidal activity was assessed using 20-h time-kill assays. Caspofungin immediately induced PI(4,5)P2 and Cdc10p colocalization to aberrant foci, a process that was highly dynamic over 3 h. PI(4,5)P2 levels increased in a dose-response manner at caspofungin concentrations of ≤4× MIC and progressively decreased at concentrations of ≥8× MIC. Caspofungin exposure resulted in broad-based mother-daughter bud necks and arrested septum-like structures, in which PI(4,5)P2 and Cdc10 colocalized. PKC-Mkc1 pathway activation was maximal within 10 min, peaked in response to caspofungin at 4× MIC, and declined at higher concentrations. The caspofungin-induced PI(4,5)P2 redistribution remained apparent in mkc1 mutants. Caspofungin exerted dose-dependent killing and paradoxical effects at ≤4× and ≥8× MIC, respectively. Fluconazole, amphotericin B, calcofluor white, and H2O2 did not impact the PI(4,5)P2 or Cdc10p distribution like caspofungin did. Caspofungin exerts rapid PI(4,5)P2-septin and PKC-Mkc1 responses that correlate with the extent of C. albicans killing, and the responses are not induced by other antifungal agents. PI(4,5)P2-septin regulation is crucial in early caspofungin responses and PKC-Mkc1 activation.

MeSH Terms
Amphotericin B/pharmacology Antifungal Agents/pharmacology Benzenesulfonates/pharmacology Candida albicans/drug effects Caspofungin Cell Cycle Proteins/genetics Cell Wall/metabolism Echinocandins/pharmacology Fluconazole/pharmacology Fungal Proteins/metabolism Green Fluorescent Proteins Hydrogen Peroxide/metabolism Lipopeptides/pharmacology Luminescent Proteins Microbial Sensitivity Tests Mitogen-Activated Protein Kinases/metabolism Phosphatidylinositol 4,5-Diphosphate/metabolism Phosphorylation Protein Domains/genetics Protein Kinase C/metabolism Septins/metabolism
Chemicals
Antifungal Agents Benzenesulfonates Cell Cycle Proteins Echinocandins Fungal Proteins Lipopeptides Luminescent Proteins Phosphatidylinositol 4,5-Diphosphate red fluorescent protein Green Fluorescent Proteins C.I. Fluorescent Brightening Agent 28 Amphotericin B Fluconazole Hydrogen Peroxide Protein Kinase C MKC1 protein, Candida albicans Mitogen-Activated Protein Kinases Septins Caspofungin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Badrane Hassan
Division of Infectious Diseases, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA [email protected].
Nguyen M Hong
Division of Infectious Diseases, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
Clancy Cornelius J
Division of Infectious Diseases, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA VA Pittsburgh Healthcare System, Pittsburgh, Pennsylvania, USA.
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Article Info
Journal
Antimicrobial agents and chemotherapy
Abbr.
Antimicrob Agents Chemother
ISSN
1098-6596
DOI
10.1128/AAC.02711-15
Email
[email protected]
Published
2016-00-00
Epub
2016-00-23
Pages
3591-600
Language
English
Region
United States
NLM ID
0315061
PMCID
PMC4879351
Subset
IM
External Links
PubMed source DOI: 10.1128/AAC.02711-15 PMC Full Text View journal details
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