Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content>
ABSTRACT Lathosterol oxidase (LSO) catalyzes the formation of the C-5–C-6 double bond in the synthesis of various types of sterols in mammals, fungi, plants, and protozoa. In Leishmania parasites, mutations in LSO or other sterol biosynthetic genes are associated with amphotericin B resistance. To i...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2020-08-01
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| Series: | mSphere |
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| Online Access: | https://journals.asm.org/doi/10.1128/mSphere.00380-20 |
| _version_ | 1818822255614885888 |
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| author | Yu Ning Cheryl Frankfater Fong-Fu Hsu Rodrigo P. Soares Camila A. Cardoso Paula M. Nogueira Noelia Marina Lander Roberto Docampo Kai Zhang |
| author_facet | Yu Ning Cheryl Frankfater Fong-Fu Hsu Rodrigo P. Soares Camila A. Cardoso Paula M. Nogueira Noelia Marina Lander Roberto Docampo Kai Zhang |
| author_sort | Yu Ning |
| collection | DOAJ |
| description | ABSTRACT Lathosterol oxidase (LSO) catalyzes the formation of the C-5–C-6 double bond in the synthesis of various types of sterols in mammals, fungi, plants, and protozoa. In Leishmania parasites, mutations in LSO or other sterol biosynthetic genes are associated with amphotericin B resistance. To investigate the biological roles of sterol C-5–C-6 desaturation, we generated an LSO-null mutant line (lso−) in Leishmania major, the causative agent for cutaneous leishmaniasis. lso− parasites lacked the ergostane-based sterols commonly found in wild-type L. major and instead accumulated equivalent sterol species without the C-5–C-6 double bond. These mutant parasites were replicative in culture and displayed heightened resistance to amphotericin B. However, they survived poorly after reaching the maximal density and were highly vulnerable to the membrane-disrupting detergent Triton X-100. In addition, lso− mutants showed defects in regulating intracellular pH and were hypersensitive to acidic conditions. They also had potential alterations in the carbohydrate composition of lipophosphoglycan, a membrane-bound virulence factor in Leishmania. All these defects in lso− were corrected upon the restoration of LSO expression. Together, these findings suggest that the C-5–C-6 double bond is vital for the structure of the sterol core, and while the loss of LSO can lead to amphotericin B resistance, it also makes Leishmania parasites vulnerable to biologically relevant stress. IMPORTANCE Sterols are essential membrane components in eukaryotes, and sterol synthesis inhibitors can have potent effects against pathogenic fungi and trypanosomatids. Understanding the roles of sterols will facilitate the development of new drugs and counter drug resistance. LSO is required for the formation of the C-5–C-6 double bond in the sterol core structure in mammals, fungi, protozoans, plants, and algae. Functions of this C-5–C-6 double bond are not well understood. In this study, we generated and characterized a lathosterol oxidase-null mutant in Leishmania major. Our data suggest that LSO is vital for the structure and membrane-stabilizing functions of leishmanial sterols. In addition, our results imply that while mutations in lathosterol oxidase can confer resistance to amphotericin B, an important antifungal and antiprotozoal agent, the alteration in sterol structure leads to significant defects in stress response that could be exploited for drug development. |
| first_indexed | 2024-12-18T23:21:11Z |
| format | Article |
| id | doaj.art-8e6b658ffd5043579442d11bcb0da4ed |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-12-18T23:21:11Z |
| publishDate | 2020-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-8e6b658ffd5043579442d11bcb0da4ed2022-12-21T20:47:57ZengAmerican Society for MicrobiologymSphere2379-50422020-08-015410.1128/mSphere.00380-20Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content>Yu Ning0Cheryl Frankfater1Fong-Fu Hsu2Rodrigo P. Soares3Camila A. Cardoso4Paula M. Nogueira5Noelia Marina Lander6Roberto Docampo7Kai Zhang8Department of Biological Sciences, Texas Tech University, Lubbock, Texas, USAMass Spectrometry Resource, Division of Endocrinology, Diabetes, Metabolism, and Lipid Research, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, USAMass Spectrometry Resource, Division of Endocrinology, Diabetes, Metabolism, and Lipid Research, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, USAFundação Oswaldo Cruz-Fiocruz, Instituto René Rachou, Belo Horizonte, Minas Gerais, BrazilFundação Oswaldo Cruz-Fiocruz, Instituto René Rachou, Belo Horizonte, Minas Gerais, BrazilFundação Oswaldo Cruz-Fiocruz, Instituto René Rachou, Belo Horizonte, Minas Gerais, BrazilCenter for Tropical and Emerging Global Diseases, University of Georgia, Athens, Georgia, USACenter for Tropical and Emerging Global Diseases, University of Georgia, Athens, Georgia, USADepartment of Biological Sciences, Texas Tech University, Lubbock, Texas, USAABSTRACT Lathosterol oxidase (LSO) catalyzes the formation of the C-5–C-6 double bond in the synthesis of various types of sterols in mammals, fungi, plants, and protozoa. In Leishmania parasites, mutations in LSO or other sterol biosynthetic genes are associated with amphotericin B resistance. To investigate the biological roles of sterol C-5–C-6 desaturation, we generated an LSO-null mutant line (lso−) in Leishmania major, the causative agent for cutaneous leishmaniasis. lso− parasites lacked the ergostane-based sterols commonly found in wild-type L. major and instead accumulated equivalent sterol species without the C-5–C-6 double bond. These mutant parasites were replicative in culture and displayed heightened resistance to amphotericin B. However, they survived poorly after reaching the maximal density and were highly vulnerable to the membrane-disrupting detergent Triton X-100. In addition, lso− mutants showed defects in regulating intracellular pH and were hypersensitive to acidic conditions. They also had potential alterations in the carbohydrate composition of lipophosphoglycan, a membrane-bound virulence factor in Leishmania. All these defects in lso− were corrected upon the restoration of LSO expression. Together, these findings suggest that the C-5–C-6 double bond is vital for the structure of the sterol core, and while the loss of LSO can lead to amphotericin B resistance, it also makes Leishmania parasites vulnerable to biologically relevant stress. IMPORTANCE Sterols are essential membrane components in eukaryotes, and sterol synthesis inhibitors can have potent effects against pathogenic fungi and trypanosomatids. Understanding the roles of sterols will facilitate the development of new drugs and counter drug resistance. LSO is required for the formation of the C-5–C-6 double bond in the sterol core structure in mammals, fungi, protozoans, plants, and algae. Functions of this C-5–C-6 double bond are not well understood. In this study, we generated and characterized a lathosterol oxidase-null mutant in Leishmania major. Our data suggest that LSO is vital for the structure and membrane-stabilizing functions of leishmanial sterols. In addition, our results imply that while mutations in lathosterol oxidase can confer resistance to amphotericin B, an important antifungal and antiprotozoal agent, the alteration in sterol structure leads to significant defects in stress response that could be exploited for drug development.https://journals.asm.org/doi/10.1128/mSphere.00380-20Leishmaniaamphotericindrug resistance mechanismslipid synthesissterolsstress response |
| spellingShingle | Yu Ning Cheryl Frankfater Fong-Fu Hsu Rodrigo P. Soares Camila A. Cardoso Paula M. Nogueira Noelia Marina Lander Roberto Docampo Kai Zhang Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content> mSphere Leishmania amphotericin drug resistance mechanisms lipid synthesis sterols stress response |
| title | Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content> |
| title_full | Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content> |
| title_fullStr | Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content> |
| title_full_unstemmed | Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content> |
| title_short | Lathosterol Oxidase (Sterol C-5 Desaturase) Deletion Confers Resistance to Amphotericin B and Sensitivity to Acidic Stress in <named-content content-type="genus-species">Leishmania major</named-content> |
| title_sort | lathosterol oxidase sterol c 5 desaturase deletion confers resistance to amphotericin b and sensitivity to acidic stress in named content content type genus species leishmania major named content |
| topic | Leishmania amphotericin drug resistance mechanisms lipid synthesis sterols stress response |
| url | https://journals.asm.org/doi/10.1128/mSphere.00380-20 |
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