A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress

ABSTRACT Cellular responses to external stress allow microorganisms to adapt to a vast array of environmental conditions, including infection sites. The molecular mechanisms behind these responses are studied to gain insight into microbial pathogenesis, which could lead to new antimicrobial therapie...

Full description

Saved in:
Bibliographic Details
Main Authors: Lukas M. du Plooy, Calla L. Telzrow, Connie B. Nichols, Corinna Probst, Natalia Castro-Lopez, Floyd L. Wormley, J. Andrew Alspaugh
Format: Article
Language:English
Published: American Society for Microbiology 2024-10-01
Series:mBio
Subjects:
Online Access:https://journals.asm.org/doi/10.1128/mbio.00981-24
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850184201427484672
author Lukas M. du Plooy
Calla L. Telzrow
Connie B. Nichols
Corinna Probst
Natalia Castro-Lopez
Floyd L. Wormley
J. Andrew Alspaugh
author_facet Lukas M. du Plooy
Calla L. Telzrow
Connie B. Nichols
Corinna Probst
Natalia Castro-Lopez
Floyd L. Wormley
J. Andrew Alspaugh
author_sort Lukas M. du Plooy
collection DOAJ
description ABSTRACT Cellular responses to external stress allow microorganisms to adapt to a vast array of environmental conditions, including infection sites. The molecular mechanisms behind these responses are studied to gain insight into microbial pathogenesis, which could lead to new antimicrobial therapies. Here, we explore a role for arrestin protein-mediated ubiquitination in stress response and pathogenesis in the pathogenic fungus Cryptococcus neoformans. In a previous study, we identified four arrestin-like proteins in C. neoformans and found that one of these is required for efficient membrane synthesis, likely by directing interaction between fatty acid synthases and the Rsp5 E3 ubiquitin ligase. Here, we further explore Cn Rsp5 function and determine that this single Ub ligase is absolutely required for pathogenesis and survival in the presence of cellular stress. Additionally, we show that a second arrestin-like protein, Ali2, similarly facilitates interaction between Rsp5 and some of its protein targets. Of the four postulated C. neoformans arrestin-like proteins, Ali2 appears to contribute the most to C. neoformans pathogenesis, likely by directing Rsp5 to pathogenesis-related ubiquitination targets. A proteomics-based differential ubiquitination screen revealed that several known cell surface proteins are ubiquitinated by Rsp5 and a subset also requires Ali2 for their ubiquitination. Rsp5-mediated ubiquitination alters the stability and the localization of these proteins. A loss of Rsp5-mediated ubiquitination results in cell wall defects that increase susceptibility to external stresses. These findings support a model in which arrestin-like proteins guide Rsp5 to ubiquitinate specific target proteins, some of which are required for survival during stress.IMPORTANCEMicrobial proteins involved in human infectious diseases often need to be modified by specific chemical additions to be fully functional. Here, we explore the role of a particular protein modification, ubiquitination, in infections due to the human fungal pathogen Cryptococcus neoformans. We identified a complex of proteins responsible for adding ubiquitin groups to fungal proteins, and this complex is required for virulence. These proteins are fungal specific and might be targets for novel anti-infection therapy.
format Article
id doaj-art-6afdb1943cbe404d9dfbeb5018096d34
institution OA Journals
issn 2150-7511
language English
publishDate 2024-10-01
publisher American Society for Microbiology
record_format Article
series mBio
spelling doaj-art-6afdb1943cbe404d9dfbeb5018096d342025-08-20T02:17:06ZengAmerican Society for MicrobiologymBio2150-75112024-10-01151010.1128/mbio.00981-24A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stressLukas M. du Plooy0Calla L. Telzrow1Connie B. Nichols2Corinna Probst3Natalia Castro-Lopez4Floyd L. Wormley5J. Andrew Alspaugh6Department of Medicine, Duke University School of Medicine, Durham, North Carolina, USADepartment of Medicine, Duke University School of Medicine, Durham, North Carolina, USADepartment of Medicine, Duke University School of Medicine, Durham, North Carolina, USADepartment of Medicine, Duke University School of Medicine, Durham, North Carolina, USADepartment of Biology, University of Texas at San Antonio, San Antonio, Texas, USADepartment of Biology, University of Texas at San Antonio, San Antonio, Texas, USADepartment of Medicine, Duke University School of Medicine, Durham, North Carolina, USAABSTRACT Cellular responses to external stress allow microorganisms to adapt to a vast array of environmental conditions, including infection sites. The molecular mechanisms behind these responses are studied to gain insight into microbial pathogenesis, which could lead to new antimicrobial therapies. Here, we explore a role for arrestin protein-mediated ubiquitination in stress response and pathogenesis in the pathogenic fungus Cryptococcus neoformans. In a previous study, we identified four arrestin-like proteins in C. neoformans and found that one of these is required for efficient membrane synthesis, likely by directing interaction between fatty acid synthases and the Rsp5 E3 ubiquitin ligase. Here, we further explore Cn Rsp5 function and determine that this single Ub ligase is absolutely required for pathogenesis and survival in the presence of cellular stress. Additionally, we show that a second arrestin-like protein, Ali2, similarly facilitates interaction between Rsp5 and some of its protein targets. Of the four postulated C. neoformans arrestin-like proteins, Ali2 appears to contribute the most to C. neoformans pathogenesis, likely by directing Rsp5 to pathogenesis-related ubiquitination targets. A proteomics-based differential ubiquitination screen revealed that several known cell surface proteins are ubiquitinated by Rsp5 and a subset also requires Ali2 for their ubiquitination. Rsp5-mediated ubiquitination alters the stability and the localization of these proteins. A loss of Rsp5-mediated ubiquitination results in cell wall defects that increase susceptibility to external stresses. These findings support a model in which arrestin-like proteins guide Rsp5 to ubiquitinate specific target proteins, some of which are required for survival during stress.IMPORTANCEMicrobial proteins involved in human infectious diseases often need to be modified by specific chemical additions to be fully functional. Here, we explore the role of a particular protein modification, ubiquitination, in infections due to the human fungal pathogen Cryptococcus neoformans. We identified a complex of proteins responsible for adding ubiquitin groups to fungal proteins, and this complex is required for virulence. These proteins are fungal specific and might be targets for novel anti-infection therapy.https://journals.asm.org/doi/10.1128/mbio.00981-24Cryptococcus neoformansarrestinE3 ubiquitin ligasefungal pathogenesis
spellingShingle Lukas M. du Plooy
Calla L. Telzrow
Connie B. Nichols
Corinna Probst
Natalia Castro-Lopez
Floyd L. Wormley
J. Andrew Alspaugh
A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
mBio
Cryptococcus neoformans
arrestin
E3 ubiquitin ligase
fungal pathogenesis
title A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
title_full A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
title_fullStr A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
title_full_unstemmed A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
title_short A fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
title_sort fungal ubiquitin ligase and arrestin binding partner contribute to pathogenesis and survival during cellular stress
topic Cryptococcus neoformans
arrestin
E3 ubiquitin ligase
fungal pathogenesis
url https://journals.asm.org/doi/10.1128/mbio.00981-24
work_keys_str_mv AT lukasmduplooy afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT callaltelzrow afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT conniebnichols afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT corinnaprobst afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT nataliacastrolopez afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT floydlwormley afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT jandrewalspaugh afungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT lukasmduplooy fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT callaltelzrow fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT conniebnichols fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT corinnaprobst fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT nataliacastrolopez fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT floydlwormley fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress
AT jandrewalspaugh fungalubiquitinligaseandarrestinbindingpartnercontributetopathogenesisandsurvivalduringcellularstress