The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells
Candida albicans causes severe invasive candidiasis. C. albicans infection requires the virulence factor candidalysin (CL) which damages target cell membranes. However, the mechanism that CL uses to permeabilize membranes is unclear. We reveal that CL forms membrane pores using a unique mechanism. U...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/75490 |
| _version_ | 1811258179625943040 |
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| author | Charles M Russell Katherine G Schaefer Andrew Dixson Amber LH Gray Robert J Pyron Daiane S Alves Nicholas Moore Elizabeth A Conley Ryan J Schuck Tommi A White Thanh D Do Gavin M King Francisco N Barrera |
| author_facet | Charles M Russell Katherine G Schaefer Andrew Dixson Amber LH Gray Robert J Pyron Daiane S Alves Nicholas Moore Elizabeth A Conley Ryan J Schuck Tommi A White Thanh D Do Gavin M King Francisco N Barrera |
| author_sort | Charles M Russell |
| collection | DOAJ |
| description | Candida albicans causes severe invasive candidiasis. C. albicans infection requires the virulence factor candidalysin (CL) which damages target cell membranes. However, the mechanism that CL uses to permeabilize membranes is unclear. We reveal that CL forms membrane pores using a unique mechanism. Unexpectedly, CL readily assembled into polymers in solution. We propose that the basic structural unit in polymer formation is a CL oligomer, which is sequentially added into a string configuration that can close into a loop. CL loops appear to spontaneously insert into the membrane to become pores. A CL mutation (G4W) inhibited the formation of polymers in solution and prevented pore formation in synthetic lipid systems. Epithelial cell studies showed that G4W CL failed to activate the danger response pathway, a hallmark of the pathogenic effect of CL. These results indicate that CL polymerization in solution is a necessary step for the damage of cellular membranes. Analysis of CL pores by atomic force microscopy revealed co-existence of simple depressions and more complex pores, which are likely formed by CL assembled in an alternate oligomer orientation. We propose that this structural rearrangement represents a maturation mechanism that stabilizes pore formation to achieve more robust cellular damage. To summarize, CL uses a previously unknown mechanism to damage membranes, whereby pre-assembly of CL loops in solution leads to formation of membrane pores. Our investigation not only unravels a new paradigm for the formation of membrane pores, but additionally identifies CL polymerization as a novel therapeutic target to treat candidiasis. |
| first_indexed | 2024-04-12T18:10:12Z |
| format | Article |
| id | doaj.art-79958a3cea334a74ad69da42c957104a |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T18:10:12Z |
| publishDate | 2022-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-79958a3cea334a74ad69da42c957104a2022-12-22T03:21:52ZengeLife Sciences Publications LtdeLife2050-084X2022-09-011110.7554/eLife.75490The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cellsCharles M Russell0https://orcid.org/0000-0003-2489-657XKatherine G Schaefer1https://orcid.org/0000-0003-2180-799XAndrew Dixson2Amber LH Gray3https://orcid.org/0000-0003-4126-868XRobert J Pyron4Daiane S Alves5https://orcid.org/0000-0001-9154-4748Nicholas Moore6Elizabeth A Conley7Ryan J Schuck8Tommi A White9Thanh D Do10https://orcid.org/0000-0002-1978-4365Gavin M King11https://orcid.org/0000-0002-5811-7012Francisco N Barrera12https://orcid.org/0000-0002-5200-7891Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesDepartment of Physics and Astronomy, University of Missouri, Columbia, United StatesDepartment of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesDepartment of Chemistry, University of Tennessee, Knoxville, United StatesDepartment of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesDepartment of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesDepartment of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesDepartment of Physics and Astronomy, University of Missouri, Columbia, United StatesDepartment of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesDepartment of Biochemistry, University of Missouri, Columbia, United States; Electron Microscopy Core, University of Missouri, Columbia, United StatesDepartment of Chemistry, University of Tennessee, Knoxville, United StatesDepartment of Physics and Astronomy, University of Missouri, Columbia, United States; Department of Biochemistry, University of Missouri, Columbia, United StatesDepartment of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, United StatesCandida albicans causes severe invasive candidiasis. C. albicans infection requires the virulence factor candidalysin (CL) which damages target cell membranes. However, the mechanism that CL uses to permeabilize membranes is unclear. We reveal that CL forms membrane pores using a unique mechanism. Unexpectedly, CL readily assembled into polymers in solution. We propose that the basic structural unit in polymer formation is a CL oligomer, which is sequentially added into a string configuration that can close into a loop. CL loops appear to spontaneously insert into the membrane to become pores. A CL mutation (G4W) inhibited the formation of polymers in solution and prevented pore formation in synthetic lipid systems. Epithelial cell studies showed that G4W CL failed to activate the danger response pathway, a hallmark of the pathogenic effect of CL. These results indicate that CL polymerization in solution is a necessary step for the damage of cellular membranes. Analysis of CL pores by atomic force microscopy revealed co-existence of simple depressions and more complex pores, which are likely formed by CL assembled in an alternate oligomer orientation. We propose that this structural rearrangement represents a maturation mechanism that stabilizes pore formation to achieve more robust cellular damage. To summarize, CL uses a previously unknown mechanism to damage membranes, whereby pre-assembly of CL loops in solution leads to formation of membrane pores. Our investigation not only unravels a new paradigm for the formation of membrane pores, but additionally identifies CL polymerization as a novel therapeutic target to treat candidiasis.https://elifesciences.org/articles/75490atomic force microscopynative mass spectrometrymass photometry |
| spellingShingle | Charles M Russell Katherine G Schaefer Andrew Dixson Amber LH Gray Robert J Pyron Daiane S Alves Nicholas Moore Elizabeth A Conley Ryan J Schuck Tommi A White Thanh D Do Gavin M King Francisco N Barrera The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells eLife atomic force microscopy native mass spectrometry mass photometry |
| title | The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells |
| title_full | The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells |
| title_fullStr | The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells |
| title_full_unstemmed | The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells |
| title_short | The Candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells |
| title_sort | candida albicans virulence factor candidalysin polymerizes in solution to form membrane pores and damage epithelial cells |
| topic | atomic force microscopy native mass spectrometry mass photometry |
| url | https://elifesciences.org/articles/75490 |
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