Skip to main content
  • ASM Journals
    • Antimicrobial Agents and Chemotheraphy
    • Applied and Environmental Mircobiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Genome Announcements
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
Advertisement

Main menu

  • Home
  • Articles
    • Latest Articles
    • Archive
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • For Authors
    • Getting Started
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Portal
  • About the Journal
    • About mSphere
    • mSphereDirect
    • Editor-in-Chief
    • Board of Editors
    • Data Policy
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • ASM Journals
    • Antimicrobial Agents and Chemotheraphy
    • Applied and Environmental Mircobiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Genome Announcements
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts

Search

  • Advanced search
mSphere

mSphere

Advanced Search

  • Home
  • Articles
    • Latest Articles
    • Archive
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • For Authors
    • Getting Started
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Portal
  • About the Journal
    • About mSphere
    • mSphereDirect
    • Editor-in-Chief
    • Board of Editors
    • Data Policy
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
Research Article | Molecular Biology and Physiology

Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A

Jinglin L. Xie, Teresa R. O’Meara, Elizabeth J. Polvi, Nicole Robbins, Leah E. Cowen
Aaron P. Mitchell, Editor
Jinglin L. Xie
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Teresa R. O’Meara
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Elizabeth J. Polvi
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicole Robbins
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Leah E. Cowen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Aaron P. Mitchell
Carnegie Mellon University
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/mSphere.00056-17
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Supplemental Material
  • FIG 1 
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG 1 

    Staurosporine induces filamentation independent of Pkc1. SN95 wild-type (WT) cells were subcultured to log phase in YPD at 30°C, YPD plus 0.5 μg/ml staurosporine at 30°C, YPD plus 10 μM geldanamycin at 30°C, Spider medium at 37°C, or 10% serum at 37°C. Cells were imaged by DIC microscopy. The scale bar indicates 20 μm. White arrows highlight representative filaments with obvious constrictions along the filament. Red arrows highlight representative filaments with a widening at the bud neck that narrows toward the tip.

  • FIG 2 
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG 2 

    Staurosporine-induced filamentation requires Cyr1 and PKA. (A) Farnesol inhibits filamentation induced by staurosporine. SN95 wild-type cells were grown to log phase at 30°C in YPD or YPD plus 0.5 μg/ml staurosporine in the absence or presence of 200 μM farnesol. Cells were imaged by DIC microscopy. The scale bar indicates 10 μm. (B) Cyr1 and PKA are the only components of the cAMP signaling pathway tested that are required for filamentation induced by staurosporine. A CAI4 wild-type strain or mutants lacking components of the cAMP signaling pathway were subcultured to stationary phase in YPD at 30°C in the presence or absence of 0.5 μg/ml staurosporine (STS). Cells were imaged by DIC microscopy. The scale bar indicates 20 μm.

  • FIG 3 
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG 3 

    Staurosporine induces a distinct gene expression program from other filament-inducing cues and promotes filamentation independent of Nrg1 degradation. (A) SN95 wild-type cells were subcultured to log phase in YPD at 30°C, YPD plus 10% serum at 37°C, YPD plus 200 mM hydroxyurea at 30°C, or YPD plus 0.5 μg/ml staurosporine at 30°C. cDNA was prepared from total RNA for qRT-PCR. The transcript levels of filament-specific transcripts (ECE1, HGC1, HWP1, and RBT1) and yeast-specific transcripts (YWP1 and NRG1) were monitored by qRT-PCR and normalized to GPD1. The fold change in gene expression under each condition relative to YPD at 30°C is plotted as the mean ± standard deviation from triplicate samples and is representative of two independent experiments. (B) Nrg1 protein persists in filaments induced by staurosporine. SN95 wild-type cells expressing native levels of HA-tagged Nrg1 were subcultured to log phase in YPD plus 10% serum at 37°C, YPD plus 200 mM hydroxyurea at 30°C, or YPD plus 0.5 μg/ml staurosporine at 30°C. Total protein was resolved by SDS-PAGE, and the blot was hybridized with anti-hemagglutinin to detect Nrg1 and anti-PSTAIRE to monitor Cdc28 as a loading control. (C) SN95 wild-type cells were grown to log phase under identical conditions as described for panel B. Cells were imaged by DIC microscopy. The scale bar indicates 10 μm.

  • FIG 4 
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG 4 

    Dibutyryl cAMP does not rescue staurosporine-induced filamentation of a cyr1Δ/cyr1Δ mutant. A CAI4 wild-type strain and a cyr1Δ/cyr1Δ mutant were grown to log phase at 37°C in YPD plus 10% serum or at 30°C in YPD plus 0.5 μg/ml staurosporine in the absence or presence of 100 mM dibutyryl cAMP, as indicated. Cells were imaged by DIC microscopy. The scale bar indicates 10 μm.

  • FIG 5 
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG 5 

    Mfg1 and Flo8 are not required for polarized growth in response to staurosporine. Strains were grown to log phase at 30°C in YPD, 30°C in YPD plus 0.5 μg/ml staurosporine, or 37°C in YPD plus 10% serum. Cells were imaged by DIC microscopy. The scale bar indicates 20 μm.

  • FIG 6 
    • Download figure
    • Open in new tab
    • Download powerpoint
    FIG 6 

    Septin ring formation and chitin-containing septum formation are aberrant in filaments formed in response to staurosporine. (A) Strains were subcultured to log phase in YPD at 30°C, YPD plus 0.5 μg/ml staurosporine at 30°C, or YPD plus 10% serum at 37°C. Shown are representative fluorescence microscopy images of SN95 wild-type cells expressing native levels of GFP-tagged Cdc10 to visualize septin (left panels) and RFP-tagged Hhf1 to visualize DNA (middle panels). The scale bar represents 10 μm. The fluorescence microscopy images were merged with DIC images (right panels). (B) Strains were subcultured to log phase in YPD at 30°C in the presence or absence of 0.5 μg/ml staurosporine. Shown are representative fluorescence microscopy images of SN95 wild-type cells expressing native levels of GFP-tagged Nop1 to visualize nuclei (middle panels) and stained with calcofluor white to visualize chitin (left panels). The scale bar represents 10 μm. The fluorescence microscopy images were merged with DIC images (right panels).

Supplemental Material

  • Figures
  • FIG S1 

    Filament-inducing cues such as Spider medium and serum require a concomitant increase in temperature to 37°C. A wild-type strain was subcultured in YPD plus 10% serum or Spider medium at 30°C or 37°C. The scale bar represents 20 μm. Download FIG S1, TIF file, 1.3 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • FIG S2 

    Staurosporine induces robust filamentation in the wild-type strain CAF2-1. CAF2-1 was subcultured in YPD or YPD plus 0.5 μg/ml staurosporine at 30°C. The scale bar represents 20 μm. Download FIG S2, TIF file, 0.2 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • FIG S3 

    Staurosporine induces filamentation independent of Nrg1 degradation. (A) Staurosporine-induced filamentation is not accompanied by Nrg1 protein degradation. SN95 wild-type cells expressing native levels of HA-tagged Nrg1 were grown in YPD plus 10% serum at 37°C, YPD plus 200 mM hydroxyurea (HU) at 30°C, or YPD plus 0.5 μg/ml staurosporine at 30°C for 5 or 65 min. Total proteins were resolved by SDS-PAGE gel, and the blot was hybridized with anti-HA to detect Nrg1 and anti-tubulin to monitor tubulin as a loading control. (B) Staurosporine induces germ tube formation at different kinetics from serum and hydroxyurea. Cells were imaged by DIC microscopy. The scale bar indicates 10 μm. Download FIG S3, TIF file, 1.3 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • TABLE S1 

    Positive regulators of filamentation from the image-based arrayed morphology screen. Download TABLE S1, XLSX file, 0.1 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • TEXT S1 

    Strain and plasmid construction. Download TEXT S1, DOCX file, 0.1 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • TABLE S2 

    Candida albicans strains used in this study. Download TABLE S2, DOCX file, 0.1 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • TABLE S3 

    Plasmids used in this study. Download TABLE S3, DOCX file, 0.1 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

  • TABLE S4 

    Oligonucleotides used in this study. Download TABLE S4, XLSX file, 0.1 MB.

    Copyright © 2017 Xie et al.

    This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

PreviousNext
Back to top
Download PDF
Citation Tools
Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A
Jinglin L. Xie, Teresa R. O’Meara, Elizabeth J. Polvi, Nicole Robbins, Leah E. Cowen
mSphere Mar 2017, 2 (2) e00056-17; DOI: 10.1128/mSphere.00056-17

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print
Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this mSphere article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A
(Your Name) has forwarded a page to you from mSphere
(Your Name) thought you would be interested in this article in mSphere.
Share
Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A
Jinglin L. Xie, Teresa R. O’Meara, Elizabeth J. Polvi, Nicole Robbins, Leah E. Cowen
mSphere Mar 2017, 2 (2) e00056-17; DOI: 10.1128/mSphere.00056-17
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • MATERIALS AND METHODS
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Candida albicans
cyclic AMP
kinase inhibitor
morphogenesis
staurosporine
virulence

Related Articles

Cited By...

About the Journal

  • About mSphere
  • mSphereDirect
  • Board of Editors
  • Data Policy
  • For Reviewers
  • For the Media
  • Embargo Policy
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ

For Authors

  • Submit a Manuscript
  • Author Warranty
  • Types of Articles
  • Getting Started
  • Contact Us
  • Ethics Portal

Follow #mSphereJ

@ASMicrobiology

       

 

Website feedback

Journals.ASM.org

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

1752 N Street N.W.
Washington DC 20036
202.737.3600
202.942.9355 fax
journals@asmusa.org

Copyright © 2018 American Society for Microbiology   eISSN 2379-5042