Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx

Introduction<p>The basal fungus Mucor lusitanicus (Mlu) is a cause of mucormycosis, with limited treatment options due to intrinsic resistance to short-tailed azoles (fluconazole, voriconazole) and echinocandins. In silico analysis identified amino-acid-substitutions Y129F and V293A in the sub...

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Autor principal: K. Rosam (22687364) (author)
Outros Autores: M. V. Keniya (22687367) (author), L.-M. Zenz (22687370) (author), C. Müller (2693209) (author), B. Sarg (22687373) (author), M. Stojanovic (15235538) (author), U. Binder (22687376) (author), B. C. Monk (22687379) (author), M. Lackner (3971021) (author)
Publicado em: 2025
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_version_ 1849927623134674944
author K. Rosam (22687364)
author2 M. V. Keniya (22687367)
L.-M. Zenz (22687370)
C. Müller (2693209)
B. Sarg (22687373)
M. Stojanovic (15235538)
U. Binder (22687376)
B. C. Monk (22687379)
M. Lackner (3971021)
author2_role author
author
author
author
author
author
author
author
author_facet K. Rosam (22687364)
M. V. Keniya (22687367)
L.-M. Zenz (22687370)
C. Müller (2693209)
B. Sarg (22687373)
M. Stojanovic (15235538)
U. Binder (22687376)
B. C. Monk (22687379)
M. Lackner (3971021)
author_role author
dc.creator.none.fl_str_mv K. Rosam (22687364)
M. V. Keniya (22687367)
L.-M. Zenz (22687370)
C. Müller (2693209)
B. Sarg (22687373)
M. Stojanovic (15235538)
U. Binder (22687376)
B. C. Monk (22687379)
M. Lackner (3971021)
dc.date.none.fl_str_mv 2025-11-26T06:33:27Z
dc.identifier.none.fl_str_mv 10.3389/fmicb.2025.1702408.s001
dc.relation.none.fl_str_mv https://figshare.com/articles/dataset/Data_Sheet_1_Intrinsic_resistance_to_short-tailed_azoles_in_the_basal_fungus_Mucor_lusitanicus_functional_analysis_of_Cyp51_isoforms_and_amino_acid_substitutions_docx/30718643
dc.rights.none.fl_str_mv CC BY 4.0
info:eu-repo/semantics/openAccess
dc.subject.none.fl_str_mv Microbiology
azole
AMR (antimicrobial resistance)
mucormycoses
mucormycetes
lanosterol 14α demethylase
Saccharomyces cerevisiae
heterologous expression
GMO (genetically modified organism)
dc.title.none.fl_str_mv Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
dc.type.none.fl_str_mv Dataset
info:eu-repo/semantics/publishedVersion
dataset
description Introduction<p>The basal fungus Mucor lusitanicus (Mlu) is a cause of mucormycosis, with limited treatment options due to intrinsic resistance to short-tailed azoles (fluconazole, voriconazole) and echinocandins. In silico analysis identified amino-acid-substitutions Y129F and V293A in the substrate-binding pocket of sterol-14α-demethylase (Cyp51, Erg11) isoform F5 as potential resistance mechanisms.</p>Methods<p>This hypothesis has been experimentally tested for M. lusitanicus by expressing its Cyp51 isoforms (MluCyp51-F1, MluCyp51-F5) and modified MluCyp51-F5 variants (F129Y, A293V, F129Y A293V) with/without their cognate NADPH-cytochrome-P450-reductase (MluCPR) in Saccharomyces cerevisiae.</p>Results<p>Strains expressing MluCyp51 isoforms +/− MluCPR showed expression levels of 38–69% compared to an overexpressed Erg11 control. Susceptibility assays confirmed that MluCyp51-F5 confers intrinsic resistance to short-tailed azoles, while substitutions F129Y, A293V, or both restored susceptibility. Growth- and susceptibility-assays revealed that the MluCyp51-F1 + CPR construct had a voriconazole MIC of 0.5 μM, while MluCyp51-F5 + CPR had a MIC of 11.5 μM, with no changes in growth rate or ergosterol composition at 1.0 μM voriconazole. Susceptibility to long-tailed azoles (e.g., posaconazole) remained unchanged for both isoforms.</p>Discussion<p>These findings demonstrate the functional expression of MluCyp51-F1 and MluCyp51-F5 isoforms in a phylogenetically distant host and confirm that conserved substitutions Y129F and V293A in MluCyp51-F5 confer intrinsic resistance to short-tailed azoles in M. lusitanicus.</p>
eu_rights_str_mv openAccess
id Manara_1fabc94533bbba0aabee99bd905e4e7b
identifier_str_mv 10.3389/fmicb.2025.1702408.s001
network_acronym_str Manara
network_name_str ManaraRepo
oai_identifier_str oai:figshare.com:article/30718643
publishDate 2025
repository.mail.fl_str_mv
repository.name.fl_str_mv
repository_id_str
rights_invalid_str_mv CC BY 4.0
spelling Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docxK. Rosam (22687364)M. V. Keniya (22687367)L.-M. Zenz (22687370)C. Müller (2693209)B. Sarg (22687373)M. Stojanovic (15235538)U. Binder (22687376)B. C. Monk (22687379)M. Lackner (3971021)MicrobiologyazoleAMR (antimicrobial resistance)mucormycosesmucormyceteslanosterol 14α demethylaseSaccharomyces cerevisiaeheterologous expressionGMO (genetically modified organism)Introduction<p>The basal fungus Mucor lusitanicus (Mlu) is a cause of mucormycosis, with limited treatment options due to intrinsic resistance to short-tailed azoles (fluconazole, voriconazole) and echinocandins. In silico analysis identified amino-acid-substitutions Y129F and V293A in the substrate-binding pocket of sterol-14α-demethylase (Cyp51, Erg11) isoform F5 as potential resistance mechanisms.</p>Methods<p>This hypothesis has been experimentally tested for M. lusitanicus by expressing its Cyp51 isoforms (MluCyp51-F1, MluCyp51-F5) and modified MluCyp51-F5 variants (F129Y, A293V, F129Y A293V) with/without their cognate NADPH-cytochrome-P450-reductase (MluCPR) in Saccharomyces cerevisiae.</p>Results<p>Strains expressing MluCyp51 isoforms +/− MluCPR showed expression levels of 38–69% compared to an overexpressed Erg11 control. Susceptibility assays confirmed that MluCyp51-F5 confers intrinsic resistance to short-tailed azoles, while substitutions F129Y, A293V, or both restored susceptibility. Growth- and susceptibility-assays revealed that the MluCyp51-F1 + CPR construct had a voriconazole MIC of 0.5 μM, while MluCyp51-F5 + CPR had a MIC of 11.5 μM, with no changes in growth rate or ergosterol composition at 1.0 μM voriconazole. Susceptibility to long-tailed azoles (e.g., posaconazole) remained unchanged for both isoforms.</p>Discussion<p>These findings demonstrate the functional expression of MluCyp51-F1 and MluCyp51-F5 isoforms in a phylogenetically distant host and confirm that conserved substitutions Y129F and V293A in MluCyp51-F5 confer intrinsic resistance to short-tailed azoles in M. lusitanicus.</p>2025-11-26T06:33:27ZDatasetinfo:eu-repo/semantics/publishedVersiondataset10.3389/fmicb.2025.1702408.s001https://figshare.com/articles/dataset/Data_Sheet_1_Intrinsic_resistance_to_short-tailed_azoles_in_the_basal_fungus_Mucor_lusitanicus_functional_analysis_of_Cyp51_isoforms_and_amino_acid_substitutions_docx/30718643CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/307186432025-11-26T06:33:27Z
spellingShingle Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
K. Rosam (22687364)
Microbiology
azole
AMR (antimicrobial resistance)
mucormycoses
mucormycetes
lanosterol 14α demethylase
Saccharomyces cerevisiae
heterologous expression
GMO (genetically modified organism)
status_str publishedVersion
title Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
title_full Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
title_fullStr Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
title_full_unstemmed Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
title_short Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
title_sort Data Sheet 1_Intrinsic resistance to short-tailed azoles in the basal fungus Mucor lusitanicus: functional analysis of Cyp51 isoforms and amino acid substitutions.docx
topic Microbiology
azole
AMR (antimicrobial resistance)
mucormycoses
mucormycetes
lanosterol 14α demethylase
Saccharomyces cerevisiae
heterologous expression
GMO (genetically modified organism)