Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection

ABSTRACT Triosephosphate isomerase (TPI) catalyzes the interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (G3P). This reaction is required for glycolysis and gluconeogenesis, and tpi has been predicted to be essential for growth of Mycobacterium tuberculosis. However...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Carolina Trujillo, Antje Blumenthal, Joeli Marrero, Kyu Y. Rhee, Dirk Schnappinger, Sabine Ehrt
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b45e48d5861d431993d2aea75d2cc850
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b45e48d5861d431993d2aea75d2cc850
record_format	dspace
spelling	oai:doaj.org-article:b45e48d5861d431993d2aea75d2cc8502021-11-15T15:45:13ZTriosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection10.1128/mBio.00085-142150-7511https://doaj.org/article/b45e48d5861d431993d2aea75d2cc8502014-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00085-14https://doaj.org/toc/2150-7511ABSTRACT Triosephosphate isomerase (TPI) catalyzes the interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (G3P). This reaction is required for glycolysis and gluconeogenesis, and tpi has been predicted to be essential for growth of Mycobacterium tuberculosis. However, when studying a conditionally regulated tpi knockdown mutant, we noticed that depletion of TPI reduced growth of M. tuberculosis in media containing a single carbon source but not in media that contained both a glycolytic and a gluconeogenic carbon source. We used such two-carbon-source media to isolate a tpi deletion (Δtpi) mutant. The Δtpi mutant did not survive with single carbon substrates but grew like wild-type (WT) M. tuberculosis in the presence of both a glycolytic and a gluconeogenic carbon source. 13C metabolite tracing revealed the accumulation of TPI substrates in Δtpi and the absence of alternative triosephosphate isomerases and metabolic bypass reactions, which confirmed the requirement of TPI for glycolysis and gluconeogenesis in M. tuberculosis. The Δtpi strain was furthermore severely attenuated in the mouse model of tuberculosis, suggesting that M. tuberculosis cannot simultaneously access sufficient quantities of glycolytic and gluconeogenic carbon substrates to establish infection in mice. IMPORTANCE The importance of central carbon metabolism for the pathogenesis of M. tuberculosis has recently been recognized, but the consequences of depleting specific metabolic enzymes remain to be identified for many enzymes. We investigated triosephosphate isomerase (TPI) because it is central to both glycolysis and gluconeogenesis and had been predicted to be essential for growth of M. tuberculosis. This work identified metabolic conditions that make TPI dispensable for M. tuberculosis growth in culture and proved that M. tuberculosis relies on a single TPI enzyme and has no metabolic bypass for the TPI-dependent interconversion of dihydroxyacetone phosphate and glyceraldehyde-3-phosphate in glycolysis and gluconeogenesis. Finally, we demonstrate that TPI is essential for growth of the pathogen in mouse lungs.Carolina TrujilloAntje BlumenthalJoeli MarreroKyu Y. RheeDirk SchnappingerSabine EhrtAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 2 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Carolina Trujillo Antje Blumenthal Joeli Marrero Kyu Y. Rhee Dirk Schnappinger Sabine Ehrt Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection
description	ABSTRACT Triosephosphate isomerase (TPI) catalyzes the interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate (G3P). This reaction is required for glycolysis and gluconeogenesis, and tpi has been predicted to be essential for growth of Mycobacterium tuberculosis. However, when studying a conditionally regulated tpi knockdown mutant, we noticed that depletion of TPI reduced growth of M. tuberculosis in media containing a single carbon source but not in media that contained both a glycolytic and a gluconeogenic carbon source. We used such two-carbon-source media to isolate a tpi deletion (Δtpi) mutant. The Δtpi mutant did not survive with single carbon substrates but grew like wild-type (WT) M. tuberculosis in the presence of both a glycolytic and a gluconeogenic carbon source. 13C metabolite tracing revealed the accumulation of TPI substrates in Δtpi and the absence of alternative triosephosphate isomerases and metabolic bypass reactions, which confirmed the requirement of TPI for glycolysis and gluconeogenesis in M. tuberculosis. The Δtpi strain was furthermore severely attenuated in the mouse model of tuberculosis, suggesting that M. tuberculosis cannot simultaneously access sufficient quantities of glycolytic and gluconeogenic carbon substrates to establish infection in mice. IMPORTANCE The importance of central carbon metabolism for the pathogenesis of M. tuberculosis has recently been recognized, but the consequences of depleting specific metabolic enzymes remain to be identified for many enzymes. We investigated triosephosphate isomerase (TPI) because it is central to both glycolysis and gluconeogenesis and had been predicted to be essential for growth of M. tuberculosis. This work identified metabolic conditions that make TPI dispensable for M. tuberculosis growth in culture and proved that M. tuberculosis relies on a single TPI enzyme and has no metabolic bypass for the TPI-dependent interconversion of dihydroxyacetone phosphate and glyceraldehyde-3-phosphate in glycolysis and gluconeogenesis. Finally, we demonstrate that TPI is essential for growth of the pathogen in mouse lungs.
format	article
author	Carolina Trujillo Antje Blumenthal Joeli Marrero Kyu Y. Rhee Dirk Schnappinger Sabine Ehrt
author_facet	Carolina Trujillo Antje Blumenthal Joeli Marrero Kyu Y. Rhee Dirk Schnappinger Sabine Ehrt
author_sort	Carolina Trujillo
title	Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection
title_short	Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection
title_full	Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection
title_fullStr	Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection
title_full_unstemmed	Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection
title_sort	triosephosphate isomerase is dispensable <italic toggle="yes">in vitro</italic> yet essential for <named-content content-type="genus-species">mycobacterium tuberculosis</named-content> to establish infection
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/b45e48d5861d431993d2aea75d2cc850
work_keys_str_mv	AT carolinatrujillo triosephosphateisomeraseisdispensableitalictoggleyesinvitroitalicyetessentialfornamedcontentcontenttypegenusspeciesmycobacteriumtuberculosisnamedcontenttoestablishinfection AT antjeblumenthal triosephosphateisomeraseisdispensableitalictoggleyesinvitroitalicyetessentialfornamedcontentcontenttypegenusspeciesmycobacteriumtuberculosisnamedcontenttoestablishinfection AT joelimarrero triosephosphateisomeraseisdispensableitalictoggleyesinvitroitalicyetessentialfornamedcontentcontenttypegenusspeciesmycobacteriumtuberculosisnamedcontenttoestablishinfection AT kyuyrhee triosephosphateisomeraseisdispensableitalictoggleyesinvitroitalicyetessentialfornamedcontentcontenttypegenusspeciesmycobacteriumtuberculosisnamedcontenttoestablishinfection AT dirkschnappinger triosephosphateisomeraseisdispensableitalictoggleyesinvitroitalicyetessentialfornamedcontentcontenttypegenusspeciesmycobacteriumtuberculosisnamedcontenttoestablishinfection AT sabineehrt triosephosphateisomeraseisdispensableitalictoggleyesinvitroitalicyetessentialfornamedcontentcontenttypegenusspeciesmycobacteriumtuberculosisnamedcontenttoestablishinfection
_version_	1718427606751117312

Triosephosphate Isomerase Is Dispensable <italic toggle="yes">In Vitro</italic> yet Essential for <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> To Establish Infection

Ejemplares similares