Intravital imaging based genetic screen reveals the transcriptional network governing Candida albicans filamentation during mammalian infection

Abstract

Candida albicans is one of the most common human fungal pathogens. C. albicans pathogenesis is tightly linked to its ability to under a morphogenetic transition from typically budding yeast to filamentous forms of hyphae and pseudohyphae. Filamentous morphogenesis is the most intensively studied C. albicans virulence traits; however, nearly all of these studies have been based on in vitro induction of filamentation. Using an intravital imaging assay of filamentation during mammalian (mouse) infection, we have screened a library of transcription factor mutants to identify those that modulate both the initiation and maintenance of filamentation in vivo. We coupled this initial screen with genetic interaction analysis and in vivo transcription profiling to characterize the transcription factor network governing filamentation in infected mammalian tissue. Three core positive (Efg1, Brg1, and Rob1) and two core negative regulators (Nrg1 and Tup1) of filament initiation were identified. No previous systematic analysis of genes affecting the elongation step has been reported and we found that large set of transcription factors affect filament elongation in vivo including four (Hms1, Lys14, War1, Dal81) with no effect on in vitro elongation. We also show that the gene targets of initiation and elongation regulators are distinct. Genetic interaction analysis of the core positive and negative regulators revealed that the master regulator Efg1 primarily functions to mediate relief of Nrg1 repression and is dispensable for expression of hypha-associated genes in vitro and in vivo. Thus, our analysis not only provide the first characterization of the transcriptional network governing C. albicans filamentation in vivo but also revealed a fundamentally new mode of function for Efg1, one of the most widely studied C. albicans transcription factors.

Data availability

All data generated or analyzed during this study are included in the manuscript, supporting files, and source data files for Fig. 1, 2, and 8 are provided. Both the raw and processed gene expression data generated by Nanostring are provided in supplementary files 3,4, 5 and 6. No sequencing data was generated in this study.

Article and author information

Author details

  1. Rohan S Wakade

    Department of Pediatrics, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Laura C Ristow

    Department of Pediatrics, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Melanie Wellington

    Department of Pediatrics, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Damian J Krysan

    Department of Pediatrics, University of Iowa, Iowa City, United States
    For correspondence
    damian-krysan@uiowa.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6330-3365

Funding

National Institute of Allergy and Infectious Diseases (R01AI133409)

  • Damian J Krysan

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols at the University of Iowa as protocol 0092064.

Reviewing Editor

  1. Arturo Casadevall, Johns Hopkins Bloomberg School of Public Health, United States

Version history

  1. Received: November 23, 2022
  2. Preprint posted: November 28, 2022 (view preprint)
  3. Accepted: February 26, 2023
  4. Accepted Manuscript published: February 27, 2023 (version 1)
  5. Version of Record published: March 8, 2023 (version 2)

Copyright

© 2023, Wakade et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Rohan S Wakade
  2. Laura C Ristow
  3. Melanie Wellington
  4. Damian J Krysan
(2023)
Intravital imaging based genetic screen reveals the transcriptional network governing Candida albicans filamentation during mammalian infection
eLife 12:e85114.
https://doi.org/10.7554/eLife.85114

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https://doi.org/10.7554/eLife.85114

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