Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis
Upon infection, Mycobacterium leprae, an obligate intracellular bacillus, induces accumulation of cholesterol-enriched lipid droplets (LDs) in Schwann cells (SCs). LDs are promptly recruited to M. leprae-containing phagosomes, and inhibition of this process decreases bacterial survival, suggesting t...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-07-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2021.709972/full |
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| author | Thabatta L. S. A. Rosa Maria Angela M. Marques Zachary DeBoard Kelly Hutchins Carlos Adriano A. Silva Christine R. Montague Tianao Yuan Julio J. Amaral Georgia C. Atella Patrícia S. Rosa Katherine A. Mattos Brian C. VanderVen Ramanuj Lahiri Nicole S. Sampson Patrick J. Brennan John T. Belisle Maria Cristina V. Pessolani Marcia Berrêdo-Pinho |
| author_facet | Thabatta L. S. A. Rosa Maria Angela M. Marques Zachary DeBoard Kelly Hutchins Carlos Adriano A. Silva Christine R. Montague Tianao Yuan Julio J. Amaral Georgia C. Atella Patrícia S. Rosa Katherine A. Mattos Brian C. VanderVen Ramanuj Lahiri Nicole S. Sampson Patrick J. Brennan John T. Belisle Maria Cristina V. Pessolani Marcia Berrêdo-Pinho |
| author_sort | Thabatta L. S. A. Rosa |
| collection | DOAJ |
| description | Upon infection, Mycobacterium leprae, an obligate intracellular bacillus, induces accumulation of cholesterol-enriched lipid droplets (LDs) in Schwann cells (SCs). LDs are promptly recruited to M. leprae-containing phagosomes, and inhibition of this process decreases bacterial survival, suggesting that LD recruitment constitutes a mechanism by which host-derived lipids are delivered to intracellular M. leprae. We previously demonstrated that M. leprae has preserved only the capacity to oxidize cholesterol to cholestenone, the first step of the normal cholesterol catabolic pathway. In this study we investigated the biochemical relevance of cholesterol oxidation on bacterial pathogenesis in SCs. Firstly, we showed that M. leprae increases the uptake of LDL-cholesterol by infected SCs. Moreover, fluorescence microscopy analysis revealed a close association between M. leprae and the internalized LDL-cholesterol within the host cell. By using Mycobacterium smegmatis mutant strains complemented with M. leprae genes, we demonstrated that ml1942 coding for 3β-hydroxysteroid dehydrogenase (3β-HSD), but not ml0389 originally annotated as cholesterol oxidase (ChoD), was responsible for the cholesterol oxidation activity detected in M. leprae. The 3β-HSD activity generates the electron donors NADH and NADPH that, respectively, fuel the M. leprae respiratory chain and provide reductive power for the biosynthesis of the dominant bacterial cell wall lipids phthiocerol dimycocerosate (PDIM) and phenolic glycolipid (PGL)-I. Inhibition of M. leprae 3β-HSD activity with the 17β-[N-(2,5-di-t-butylphenyl)carbamoyl]-6-azaandrost-4-en-3one (compound 1), decreased bacterial intracellular survival in SCs. In conclusion, our findings confirm the accumulation of cholesterol in infected SCs and its potential delivery to the intracellular bacterium. Furthermore, we provide strong evidence that cholesterol oxidation is an essential catabolic pathway for M. leprae pathogenicity and point to 3β-HSD as a prime drug target that may be used in combination with current multidrug regimens to shorten leprosy treatment and ameliorate nerve damage. |
| first_indexed | 2024-12-21T16:22:32Z |
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| id | doaj.art-8febf0d55d394f6994a5972e33c674cd |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-21T16:22:32Z |
| publishDate | 2021-07-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-8febf0d55d394f6994a5972e33c674cd2022-12-21T18:57:32ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882021-07-011110.3389/fcimb.2021.709972709972Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP SynthesisThabatta L. S. A. Rosa0Maria Angela M. Marques1Zachary DeBoard2Kelly Hutchins3Carlos Adriano A. Silva4Christine R. Montague5Tianao Yuan6Julio J. Amaral7Georgia C. Atella8Patrícia S. Rosa9Katherine A. Mattos10Brian C. VanderVen11Ramanuj Lahiri12Nicole S. Sampson13Patrick J. Brennan14John T. Belisle15Maria Cristina V. Pessolani16Marcia Berrêdo-Pinho17Laboratório de Microbiologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilDepartment of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United StatesDepartment of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United StatesDepartment of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United StatesLaboratório de Microbiologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilDepartment of Microbiology and Immunology, Cornell University, Ithaca, NY, United StatesDepartment of Chemistry, Stony Brook University, Stony Brook, NY, United StatesLaboratório de Química Biológica, Diretoria de Metrologia Aplicada às Ciências da Vida, Instituto Nacional de Metrologia, Qualidade e Tecnologia, Rio de Janeiro, BrazilLaboratório de Bioquímica de Lipídeos e Lipoproteínas, Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, BrazilDivisão de Pesquisa e Ensino, Instituto Lauro de Souza Lima, Bauru, BrazilDepartmento de Controle de Qualidade, Instituto de Tecnologia em Imunobiológicos, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilDepartment of Microbiology and Immunology, Cornell University, Ithaca, NY, United StatesDepartment of Health and Human Services, Health Resources and Services Administration, Healthcare Systems Bureau, National Hansen’s Disease Programs, Baton Rouge, LA, United StatesDepartment of Chemistry, Stony Brook University, Stony Brook, NY, United StatesDepartment of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United StatesDepartment of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, United StatesLaboratório de Microbiologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilLaboratório de Microbiologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilUpon infection, Mycobacterium leprae, an obligate intracellular bacillus, induces accumulation of cholesterol-enriched lipid droplets (LDs) in Schwann cells (SCs). LDs are promptly recruited to M. leprae-containing phagosomes, and inhibition of this process decreases bacterial survival, suggesting that LD recruitment constitutes a mechanism by which host-derived lipids are delivered to intracellular M. leprae. We previously demonstrated that M. leprae has preserved only the capacity to oxidize cholesterol to cholestenone, the first step of the normal cholesterol catabolic pathway. In this study we investigated the biochemical relevance of cholesterol oxidation on bacterial pathogenesis in SCs. Firstly, we showed that M. leprae increases the uptake of LDL-cholesterol by infected SCs. Moreover, fluorescence microscopy analysis revealed a close association between M. leprae and the internalized LDL-cholesterol within the host cell. By using Mycobacterium smegmatis mutant strains complemented with M. leprae genes, we demonstrated that ml1942 coding for 3β-hydroxysteroid dehydrogenase (3β-HSD), but not ml0389 originally annotated as cholesterol oxidase (ChoD), was responsible for the cholesterol oxidation activity detected in M. leprae. The 3β-HSD activity generates the electron donors NADH and NADPH that, respectively, fuel the M. leprae respiratory chain and provide reductive power for the biosynthesis of the dominant bacterial cell wall lipids phthiocerol dimycocerosate (PDIM) and phenolic glycolipid (PGL)-I. Inhibition of M. leprae 3β-HSD activity with the 17β-[N-(2,5-di-t-butylphenyl)carbamoyl]-6-azaandrost-4-en-3one (compound 1), decreased bacterial intracellular survival in SCs. In conclusion, our findings confirm the accumulation of cholesterol in infected SCs and its potential delivery to the intracellular bacterium. Furthermore, we provide strong evidence that cholesterol oxidation is an essential catabolic pathway for M. leprae pathogenicity and point to 3β-HSD as a prime drug target that may be used in combination with current multidrug regimens to shorten leprosy treatment and ameliorate nerve damage.https://www.frontiersin.org/articles/10.3389/fcimb.2021.709972/fullMycobacterium lepraecholesterolcholestenonePGL-IPDIM3β-HSD |
| spellingShingle | Thabatta L. S. A. Rosa Maria Angela M. Marques Zachary DeBoard Kelly Hutchins Carlos Adriano A. Silva Christine R. Montague Tianao Yuan Julio J. Amaral Georgia C. Atella Patrícia S. Rosa Katherine A. Mattos Brian C. VanderVen Ramanuj Lahiri Nicole S. Sampson Patrick J. Brennan John T. Belisle Maria Cristina V. Pessolani Marcia Berrêdo-Pinho Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis Frontiers in Cellular and Infection Microbiology Mycobacterium leprae cholesterol cholestenone PGL-I PDIM 3β-HSD |
| title | Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis |
| title_full | Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis |
| title_fullStr | Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis |
| title_full_unstemmed | Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis |
| title_short | Reductive Power Generated by Mycobacterium leprae Through Cholesterol Oxidation Contributes to Lipid and ATP Synthesis |
| title_sort | reductive power generated by mycobacterium leprae through cholesterol oxidation contributes to lipid and atp synthesis |
| topic | Mycobacterium leprae cholesterol cholestenone PGL-I PDIM 3β-HSD |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2021.709972/full |
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