Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.

Mycobacterium tuberculosis (Mtb) releases the unusual terpene nucleoside 1-tuberculosinyladenosine (1-TbAd) to block lysosomal function and promote survival in human macrophages. Using conventional approaches, we found that genes Rv3377c and Rv3378c, but not Rv3376, were necessary for 1-TbAd biosynt...

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Main Authors: Jacob A Mayfield, Sahadevan Raman, Alexandrea K Ramnarine, Vivek K Mishra, Annie D Huang, Sandrine Dudoit, Jeffrey Buter, Tan-Yun Cheng, David C Young, Yashodhan M Nair, Isobel G Ouellet, Braden T Griebel, Shuyi Ma, David R Sherman, Ludovic Mallet, Kyu Y Rhee, Adriaan J Minnaard, D Branch Moody
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2024-09-01
Series:PLoS Biology
Online Access:https://doi.org/10.1371/journal.pbio.3002813
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author Jacob A Mayfield
Sahadevan Raman
Alexandrea K Ramnarine
Vivek K Mishra
Annie D Huang
Sandrine Dudoit
Jeffrey Buter
Tan-Yun Cheng
David C Young
Yashodhan M Nair
Isobel G Ouellet
Braden T Griebel
Shuyi Ma
David R Sherman
Ludovic Mallet
Kyu Y Rhee
Adriaan J Minnaard
D Branch Moody
author_facet Jacob A Mayfield
Sahadevan Raman
Alexandrea K Ramnarine
Vivek K Mishra
Annie D Huang
Sandrine Dudoit
Jeffrey Buter
Tan-Yun Cheng
David C Young
Yashodhan M Nair
Isobel G Ouellet
Braden T Griebel
Shuyi Ma
David R Sherman
Ludovic Mallet
Kyu Y Rhee
Adriaan J Minnaard
D Branch Moody
author_sort Jacob A Mayfield
collection DOAJ
description Mycobacterium tuberculosis (Mtb) releases the unusual terpene nucleoside 1-tuberculosinyladenosine (1-TbAd) to block lysosomal function and promote survival in human macrophages. Using conventional approaches, we found that genes Rv3377c and Rv3378c, but not Rv3376, were necessary for 1-TbAd biosynthesis. Here, we introduce linear models for mass spectrometry (limms) software as a next-generation lipidomics tool to study the essential functions of lipid biosynthetic enzymes on a whole-cell basis. Using limms, whole-cell lipid profiles deepened the phenotypic landscape of comparative mass spectrometry experiments and identified a large family of approximately 100 terpene nucleoside metabolites downstream of Rv3378c. We validated the identity of previously unknown adenine-, adenosine-, and lipid-modified tuberculosinol-containing molecules using synthetic chemistry and collisional mass spectrometry, including comprehensive profiling of bacterial lipids that fragment to adenine. We tracked terpene nucleoside genotypes and lipid phenotypes among Mycobacterium tuberculosis complex (MTC) species that did or did not evolve to productively infect either human or nonhuman mammals. Although 1-TbAd biosynthesis genes were thought to be restricted to the MTC, we identified the locus in unexpected species outside the MTC. Sequence analysis of the locus showed nucleotide usage characteristic of plasmids from plant-associated bacteria, clarifying the origin and timing of horizontal gene transfer to a pre-MTC progenitor. The data demonstrated correlation between high level terpene nucleoside biosynthesis and mycobacterial competence for human infection, and 2 mechanisms of 1-TbAd biosynthesis loss. Overall, the selective gain and evolutionary retention of tuberculosinyl metabolites in modern species that cause human TB suggest a role in human TB disease, and the newly discovered molecules represent candidate disease-specific biomarkers.
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spelling doaj-art-ebb47c13ec454405ad6a796ddd4a62ef2025-08-20T02:46:54ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852024-09-01229e300281310.1371/journal.pbio.3002813Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.Jacob A MayfieldSahadevan RamanAlexandrea K RamnarineVivek K MishraAnnie D HuangSandrine DudoitJeffrey ButerTan-Yun ChengDavid C YoungYashodhan M NairIsobel G OuelletBraden T GriebelShuyi MaDavid R ShermanLudovic MalletKyu Y RheeAdriaan J MinnaardD Branch MoodyMycobacterium tuberculosis (Mtb) releases the unusual terpene nucleoside 1-tuberculosinyladenosine (1-TbAd) to block lysosomal function and promote survival in human macrophages. Using conventional approaches, we found that genes Rv3377c and Rv3378c, but not Rv3376, were necessary for 1-TbAd biosynthesis. Here, we introduce linear models for mass spectrometry (limms) software as a next-generation lipidomics tool to study the essential functions of lipid biosynthetic enzymes on a whole-cell basis. Using limms, whole-cell lipid profiles deepened the phenotypic landscape of comparative mass spectrometry experiments and identified a large family of approximately 100 terpene nucleoside metabolites downstream of Rv3378c. We validated the identity of previously unknown adenine-, adenosine-, and lipid-modified tuberculosinol-containing molecules using synthetic chemistry and collisional mass spectrometry, including comprehensive profiling of bacterial lipids that fragment to adenine. We tracked terpene nucleoside genotypes and lipid phenotypes among Mycobacterium tuberculosis complex (MTC) species that did or did not evolve to productively infect either human or nonhuman mammals. Although 1-TbAd biosynthesis genes were thought to be restricted to the MTC, we identified the locus in unexpected species outside the MTC. Sequence analysis of the locus showed nucleotide usage characteristic of plasmids from plant-associated bacteria, clarifying the origin and timing of horizontal gene transfer to a pre-MTC progenitor. The data demonstrated correlation between high level terpene nucleoside biosynthesis and mycobacterial competence for human infection, and 2 mechanisms of 1-TbAd biosynthesis loss. Overall, the selective gain and evolutionary retention of tuberculosinyl metabolites in modern species that cause human TB suggest a role in human TB disease, and the newly discovered molecules represent candidate disease-specific biomarkers.https://doi.org/10.1371/journal.pbio.3002813
spellingShingle Jacob A Mayfield
Sahadevan Raman
Alexandrea K Ramnarine
Vivek K Mishra
Annie D Huang
Sandrine Dudoit
Jeffrey Buter
Tan-Yun Cheng
David C Young
Yashodhan M Nair
Isobel G Ouellet
Braden T Griebel
Shuyi Ma
David R Sherman
Ludovic Mallet
Kyu Y Rhee
Adriaan J Minnaard
D Branch Moody
Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.
PLoS Biology
title Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.
title_full Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.
title_fullStr Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.
title_full_unstemmed Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.
title_short Mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides.
title_sort mycobacteria that cause tuberculosis have retained ancestrally acquired genes for the biosynthesis of chemically diverse terpene nucleosides
url https://doi.org/10.1371/journal.pbio.3002813
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