Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>

ABSTRACT While research has identified an important contribution for metals, such as iron, in microbial pathogenesis, the roles of other transition metals, such as copper, remain mostly unknown. Recent evidence points to a requirement for copper homeostasis in the virulence of Cryptococcus neoforman...

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Autores principales:	Scott R. Waterman, Yoon-Dong Park, Meera Raja, Jin Qiu, Dima A. Hammoud, Thomas V. O’Halloran, Peter R. Williamson
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2012
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2cf5dd808ac14b94bc81076eb891abce
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spelling	oai:doaj.org-article:2cf5dd808ac14b94bc81076eb891abce2021-11-15T15:39:12ZRole of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>10.1128/mBio.00285-122150-7511https://doaj.org/article/2cf5dd808ac14b94bc81076eb891abce2012-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00285-12https://doaj.org/toc/2150-7511ABSTRACT While research has identified an important contribution for metals, such as iron, in microbial pathogenesis, the roles of other transition metals, such as copper, remain mostly unknown. Recent evidence points to a requirement for copper homeostasis in the virulence of Cryptococcus neoformans based on a role for a CUF1 copper regulatory factor in mouse models and in a human patient cohort. C. neoformans is an important fungal pathogen that results in an estimated 600,000 AIDS-related deaths yearly. In the present studies, we found that a C. neoformans mutant lacking the CUF1-dependent copper transporter, CTR4, grows normally in rich medium at 37°C but has reduced survival in macrophages and attenuated virulence in a mouse model. This reduced survival and virulence were traced to a growth defect under nutrient-restricted conditions. Expression studies using a full-length CTR4-fluorescent fusion reporter construct demonstrated robust expression in macrophages, brain, and lung, the latter shown by ex vivo fluorescent imaging. Inductively coupled mass spectroscopy (ICP-MS) was used to probe the copper quota of fungal cells grown in defined medium and recovered from brain, which suggested a role for a copper-protective function of CTR4 in combination with cell metallothioneins under copper-replete conditions. In summary, these data suggest a role for CTR4 in copper-related homeostasis and subsequently in fungal virulence. IMPORTANCE Crytococcus neoformans is a significant global fungal pathogen, and copper homeostasis is a relatively unexplored aspect of microbial pathogenesis that could lead to novel therapeutics. Previous studies correlated expression levels of a Ctr4 copper transporter to development of meningoencephalitis in a patient cohort of solid-organ transplants, but a direct role for Ctr4 in mammalian pathogenesis has not been demonstrated. The present studies utilize a Δctr4 mutant strain which revealed an important role for CTR4 in C. neoformans infections in mice and relate the gene product to homeostatic control of copper and growth under nutrient-restricted conditions. Robust expression levels of CTR4 during fungal infection were exploited to demonstrate expression and lung cryptococcal disease using ex vivo fluorescence imaging. In summary, these studies are the first to directly demonstrate a role for a copper transporter in fungal disease and provide an ex vivo imaging tool for further study of cryptococcal gene expression and pathogenesis.Scott R. WatermanYoon-Dong ParkMeera RajaJin QiuDima A. HammoudThomas V. O’HalloranPeter R. WilliamsonAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 3, Iss 5 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Scott R. Waterman Yoon-Dong Park Meera Raja Jin Qiu Dima A. Hammoud Thomas V. O’Halloran Peter R. Williamson Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>
description	ABSTRACT While research has identified an important contribution for metals, such as iron, in microbial pathogenesis, the roles of other transition metals, such as copper, remain mostly unknown. Recent evidence points to a requirement for copper homeostasis in the virulence of Cryptococcus neoformans based on a role for a CUF1 copper regulatory factor in mouse models and in a human patient cohort. C. neoformans is an important fungal pathogen that results in an estimated 600,000 AIDS-related deaths yearly. In the present studies, we found that a C. neoformans mutant lacking the CUF1-dependent copper transporter, CTR4, grows normally in rich medium at 37°C but has reduced survival in macrophages and attenuated virulence in a mouse model. This reduced survival and virulence were traced to a growth defect under nutrient-restricted conditions. Expression studies using a full-length CTR4-fluorescent fusion reporter construct demonstrated robust expression in macrophages, brain, and lung, the latter shown by ex vivo fluorescent imaging. Inductively coupled mass spectroscopy (ICP-MS) was used to probe the copper quota of fungal cells grown in defined medium and recovered from brain, which suggested a role for a copper-protective function of CTR4 in combination with cell metallothioneins under copper-replete conditions. In summary, these data suggest a role for CTR4 in copper-related homeostasis and subsequently in fungal virulence. IMPORTANCE Crytococcus neoformans is a significant global fungal pathogen, and copper homeostasis is a relatively unexplored aspect of microbial pathogenesis that could lead to novel therapeutics. Previous studies correlated expression levels of a Ctr4 copper transporter to development of meningoencephalitis in a patient cohort of solid-organ transplants, but a direct role for Ctr4 in mammalian pathogenesis has not been demonstrated. The present studies utilize a Δctr4 mutant strain which revealed an important role for CTR4 in C. neoformans infections in mice and relate the gene product to homeostatic control of copper and growth under nutrient-restricted conditions. Robust expression levels of CTR4 during fungal infection were exploited to demonstrate expression and lung cryptococcal disease using ex vivo fluorescence imaging. In summary, these studies are the first to directly demonstrate a role for a copper transporter in fungal disease and provide an ex vivo imaging tool for further study of cryptococcal gene expression and pathogenesis.
format	article
author	Scott R. Waterman Yoon-Dong Park Meera Raja Jin Qiu Dima A. Hammoud Thomas V. O’Halloran Peter R. Williamson
author_facet	Scott R. Waterman Yoon-Dong Park Meera Raja Jin Qiu Dima A. Hammoud Thomas V. O’Halloran Peter R. Williamson
author_sort	Scott R. Waterman
title	Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>
title_short	Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>
title_full	Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>
title_fullStr	Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>
title_full_unstemmed	Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>
title_sort	role of <italic toggle="yes">ctr4</italic> in the virulence of <named-content content-type="genus-species">cryptococcus neoformans</named-content>
publisher	American Society for Microbiology
publishDate	2012
url	https://doaj.org/article/2cf5dd808ac14b94bc81076eb891abce
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Role of <italic toggle="yes">CTR4</italic> in the Virulence of <named-content content-type="genus-species">Cryptococcus neoformans</named-content>

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