Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani

Jessica L. Martin, Phillip Yates, Radika Soysa, Joshua F. Alfaro, Feng Yang, Kristin E. Burnum-Johnson, Vladislav A. Petyuk, Karl K. Weitz, David G. Camp, Richard D. Smith, Phillip Wilmarth, Larry David, Gowthaman Ramasamy, Peter J. Myler, Nicola Carter

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The ability of Leishmania to survive in their insect or mammalian host is dependent upon an ability to sense and adapt to changes in the microenvironment. However, little is known about the molecular mechanisms underlying the parasite response to environmental changes, such as nutrient availability. To elucidate nutrient stress response pathways in Leishmania donovani, we have used purine starvation as the paradigm. The salvage of purines from the host milieu is obligatory for parasite replication; nevertheless, purine-starved parasites can persist in culture without supplementary purine for over three months, indicating that the response to purine starvation is robust and engenders parasite survival under conditions of extreme scarcity. To understand metabolic reprogramming during purine starvation we have employed global approaches. Whole proteome comparisons between purine-starved and purine-replete parasites over a 6-48 h span have revealed a temporal and coordinated response to purine starvation. Purine transporters and enzymes involved in acquisition at the cell surface are upregulated within a few hours of purine removal from the media, while other key purine salvage components are upregulated later in the time-course and more modestly. After 48 h, the proteome of purine-starved parasites is extensively remodeled and adaptations to purine stress appear tailored to deal with both purine deprivation and general stress. To probe the molecular mechanisms affecting proteome remodeling in response to purine starvation, comparative RNA-seq analyses, qRT-PCR, and luciferase reporter assays were performed on purine-starved versus purine-replete parasites. While the regulation of a minority of proteins tracked with changes at the mRNA level, for many regulated proteins it appears that proteome remodeling during purine stress occurs primarily via translational and/or post-translational mechanisms.

Original languageEnglish (US)
Article numbere1003938
JournalPLoS Pathogens
Volume10
Issue number2
DOIs
StatePublished - 2014

Fingerprint

Leishmania donovani
Parasites
Starvation
Proteome
purine
Molecular Probes
Food
Purines

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Martin, J. L., Yates, P., Soysa, R., Alfaro, J. F., Yang, F., Burnum-Johnson, K. E., ... Carter, N. (2014). Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani. PLoS Pathogens, 10(2), [e1003938]. https://doi.org/10.1371/journal.ppat.1003938

Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani. / Martin, Jessica L.; Yates, Phillip; Soysa, Radika; Alfaro, Joshua F.; Yang, Feng; Burnum-Johnson, Kristin E.; Petyuk, Vladislav A.; Weitz, Karl K.; Camp, David G.; Smith, Richard D.; Wilmarth, Phillip; David, Larry; Ramasamy, Gowthaman; Myler, Peter J.; Carter, Nicola.

In: PLoS Pathogens, Vol. 10, No. 2, e1003938, 2014.

Research output: Contribution to journalArticle

Martin, JL, Yates, P, Soysa, R, Alfaro, JF, Yang, F, Burnum-Johnson, KE, Petyuk, VA, Weitz, KK, Camp, DG, Smith, RD, Wilmarth, P, David, L, Ramasamy, G, Myler, PJ & Carter, N 2014, 'Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani', PLoS Pathogens, vol. 10, no. 2, e1003938. https://doi.org/10.1371/journal.ppat.1003938
Martin, Jessica L. ; Yates, Phillip ; Soysa, Radika ; Alfaro, Joshua F. ; Yang, Feng ; Burnum-Johnson, Kristin E. ; Petyuk, Vladislav A. ; Weitz, Karl K. ; Camp, David G. ; Smith, Richard D. ; Wilmarth, Phillip ; David, Larry ; Ramasamy, Gowthaman ; Myler, Peter J. ; Carter, Nicola. / Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani. In: PLoS Pathogens. 2014 ; Vol. 10, No. 2.
@article{612a379228174fa5bb0155a6e930855a,
title = "Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani",
abstract = "The ability of Leishmania to survive in their insect or mammalian host is dependent upon an ability to sense and adapt to changes in the microenvironment. However, little is known about the molecular mechanisms underlying the parasite response to environmental changes, such as nutrient availability. To elucidate nutrient stress response pathways in Leishmania donovani, we have used purine starvation as the paradigm. The salvage of purines from the host milieu is obligatory for parasite replication; nevertheless, purine-starved parasites can persist in culture without supplementary purine for over three months, indicating that the response to purine starvation is robust and engenders parasite survival under conditions of extreme scarcity. To understand metabolic reprogramming during purine starvation we have employed global approaches. Whole proteome comparisons between purine-starved and purine-replete parasites over a 6-48 h span have revealed a temporal and coordinated response to purine starvation. Purine transporters and enzymes involved in acquisition at the cell surface are upregulated within a few hours of purine removal from the media, while other key purine salvage components are upregulated later in the time-course and more modestly. After 48 h, the proteome of purine-starved parasites is extensively remodeled and adaptations to purine stress appear tailored to deal with both purine deprivation and general stress. To probe the molecular mechanisms affecting proteome remodeling in response to purine starvation, comparative RNA-seq analyses, qRT-PCR, and luciferase reporter assays were performed on purine-starved versus purine-replete parasites. While the regulation of a minority of proteins tracked with changes at the mRNA level, for many regulated proteins it appears that proteome remodeling during purine stress occurs primarily via translational and/or post-translational mechanisms.",
author = "Martin, {Jessica L.} and Phillip Yates and Radika Soysa and Alfaro, {Joshua F.} and Feng Yang and Burnum-Johnson, {Kristin E.} and Petyuk, {Vladislav A.} and Weitz, {Karl K.} and Camp, {David G.} and Smith, {Richard D.} and Phillip Wilmarth and Larry David and Gowthaman Ramasamy and Myler, {Peter J.} and Nicola Carter",
year = "2014",
doi = "10.1371/journal.ppat.1003938",
language = "English (US)",
volume = "10",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "2",

}

TY - JOUR

T1 - Metabolic Reprogramming during Purine Stress in the Protozoan Pathogen Leishmania donovani

AU - Martin, Jessica L.

AU - Yates, Phillip

AU - Soysa, Radika

AU - Alfaro, Joshua F.

AU - Yang, Feng

AU - Burnum-Johnson, Kristin E.

AU - Petyuk, Vladislav A.

AU - Weitz, Karl K.

AU - Camp, David G.

AU - Smith, Richard D.

AU - Wilmarth, Phillip

AU - David, Larry

AU - Ramasamy, Gowthaman

AU - Myler, Peter J.

AU - Carter, Nicola

PY - 2014

Y1 - 2014

N2 - The ability of Leishmania to survive in their insect or mammalian host is dependent upon an ability to sense and adapt to changes in the microenvironment. However, little is known about the molecular mechanisms underlying the parasite response to environmental changes, such as nutrient availability. To elucidate nutrient stress response pathways in Leishmania donovani, we have used purine starvation as the paradigm. The salvage of purines from the host milieu is obligatory for parasite replication; nevertheless, purine-starved parasites can persist in culture without supplementary purine for over three months, indicating that the response to purine starvation is robust and engenders parasite survival under conditions of extreme scarcity. To understand metabolic reprogramming during purine starvation we have employed global approaches. Whole proteome comparisons between purine-starved and purine-replete parasites over a 6-48 h span have revealed a temporal and coordinated response to purine starvation. Purine transporters and enzymes involved in acquisition at the cell surface are upregulated within a few hours of purine removal from the media, while other key purine salvage components are upregulated later in the time-course and more modestly. After 48 h, the proteome of purine-starved parasites is extensively remodeled and adaptations to purine stress appear tailored to deal with both purine deprivation and general stress. To probe the molecular mechanisms affecting proteome remodeling in response to purine starvation, comparative RNA-seq analyses, qRT-PCR, and luciferase reporter assays were performed on purine-starved versus purine-replete parasites. While the regulation of a minority of proteins tracked with changes at the mRNA level, for many regulated proteins it appears that proteome remodeling during purine stress occurs primarily via translational and/or post-translational mechanisms.

AB - The ability of Leishmania to survive in their insect or mammalian host is dependent upon an ability to sense and adapt to changes in the microenvironment. However, little is known about the molecular mechanisms underlying the parasite response to environmental changes, such as nutrient availability. To elucidate nutrient stress response pathways in Leishmania donovani, we have used purine starvation as the paradigm. The salvage of purines from the host milieu is obligatory for parasite replication; nevertheless, purine-starved parasites can persist in culture without supplementary purine for over three months, indicating that the response to purine starvation is robust and engenders parasite survival under conditions of extreme scarcity. To understand metabolic reprogramming during purine starvation we have employed global approaches. Whole proteome comparisons between purine-starved and purine-replete parasites over a 6-48 h span have revealed a temporal and coordinated response to purine starvation. Purine transporters and enzymes involved in acquisition at the cell surface are upregulated within a few hours of purine removal from the media, while other key purine salvage components are upregulated later in the time-course and more modestly. After 48 h, the proteome of purine-starved parasites is extensively remodeled and adaptations to purine stress appear tailored to deal with both purine deprivation and general stress. To probe the molecular mechanisms affecting proteome remodeling in response to purine starvation, comparative RNA-seq analyses, qRT-PCR, and luciferase reporter assays were performed on purine-starved versus purine-replete parasites. While the regulation of a minority of proteins tracked with changes at the mRNA level, for many regulated proteins it appears that proteome remodeling during purine stress occurs primarily via translational and/or post-translational mechanisms.

UR - http://www.scopus.com/inward/record.url?scp=84895744136&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84895744136&partnerID=8YFLogxK

U2 - 10.1371/journal.ppat.1003938

DO - 10.1371/journal.ppat.1003938

M3 - Article

VL - 10

JO - PLoS Pathogens

JF - PLoS Pathogens

SN - 1553-7366

IS - 2

M1 - e1003938

ER -