Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes

Jan M. Boitz, Caslin A. Gilroy, Tamara D. Olenyik, Dustin Paradis, Jasmine Perdeh, Kristie Dearman, Madison J. Davis, Phillip Yates, Yuexin Li, Michael Riscoe, Buddy Ullman, Sigrid C. Roberts

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Studies of Leishmania donovani have shown that both ornithine decarboxylase and spermidine synthase, two enzymes of the polyamine biosynthetic pathway, are critical for promastigote proliferation and required for maximum infection in mice. However, the importance of arginase (ARG), the first enzyme of the polyamine pathway in Leishmania, has not been analyzed in L. donovani. To test ARG function in intact parasites, we generated Δarg null mutants in L. donovani and evaluated their ability to proliferate in vitro and trigger infections in mice. The Δarg knockout was incapable of growth in the absence of polyamine supplementation, but the auxotrophic phenotype could be bypassed by addition of either millimolar concentrations of ornithine or micromolar concentrations of putrescine or by complementation with either glycosomal or cytosolic versions of ARG. Spermidine supplementation of the medium did not circumvent the polyamine auxotrophy of the Δarg line. Although ARG was found to be essential for ornithine and polyamine synthesis, ornithine decarboxylase appeared to be the rate-limiting enzyme for polyamine production. Mouse infectivity studies revealed that the Δarg lesion reduced parasite burdens in livers by an order of magnitude but had little impact on the numbers of parasites recovered from spleens. Thus, ARG is essential for proliferation of promastigotes but not intracellular amastigotes. Coupled with previous studies, these data support a model in which L. donovani amastigotes readily salvage ornithine and have some access to host spermidine pools, while host putrescine appears to be unavailable for salvage by the parasite.

Original languageEnglish (US)
Article numbere00554-16
JournalInfection and Immunity
Volume85
Issue number1
DOIs
StatePublished - 2017

Fingerprint

Leishmania donovani
Arginase
Polyamines
Ornithine
Parasites
Ornithine Decarboxylase
Putrescine
Spermidine
Spermidine Synthase
Enzymes
Leishmania
Biosynthetic Pathways
Infection
Spleen
Phenotype
Liver
Growth

Keywords

  • Arginase
  • Leishmania
  • Polyamines

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

Cite this

Boitz, J. M., Gilroy, C. A., Olenyik, T. D., Paradis, D., Perdeh, J., Dearman, K., ... Roberts, S. C. (2017). Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes. Infection and Immunity, 85(1), [e00554-16]. https://doi.org/10.1128/IAI.00554-16

Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes. / Boitz, Jan M.; Gilroy, Caslin A.; Olenyik, Tamara D.; Paradis, Dustin; Perdeh, Jasmine; Dearman, Kristie; Davis, Madison J.; Yates, Phillip; Li, Yuexin; Riscoe, Michael; Ullman, Buddy; Roberts, Sigrid C.

In: Infection and Immunity, Vol. 85, No. 1, e00554-16, 2017.

Research output: Contribution to journalArticle

Boitz, JM, Gilroy, CA, Olenyik, TD, Paradis, D, Perdeh, J, Dearman, K, Davis, MJ, Yates, P, Li, Y, Riscoe, M, Ullman, B & Roberts, SC 2017, 'Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes', Infection and Immunity, vol. 85, no. 1, e00554-16. https://doi.org/10.1128/IAI.00554-16
Boitz, Jan M. ; Gilroy, Caslin A. ; Olenyik, Tamara D. ; Paradis, Dustin ; Perdeh, Jasmine ; Dearman, Kristie ; Davis, Madison J. ; Yates, Phillip ; Li, Yuexin ; Riscoe, Michael ; Ullman, Buddy ; Roberts, Sigrid C. / Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes. In: Infection and Immunity. 2017 ; Vol. 85, No. 1.
@article{b779defeb6cc421eb155b5e151d27799,
title = "Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes",
abstract = "Studies of Leishmania donovani have shown that both ornithine decarboxylase and spermidine synthase, two enzymes of the polyamine biosynthetic pathway, are critical for promastigote proliferation and required for maximum infection in mice. However, the importance of arginase (ARG), the first enzyme of the polyamine pathway in Leishmania, has not been analyzed in L. donovani. To test ARG function in intact parasites, we generated Δarg null mutants in L. donovani and evaluated their ability to proliferate in vitro and trigger infections in mice. The Δarg knockout was incapable of growth in the absence of polyamine supplementation, but the auxotrophic phenotype could be bypassed by addition of either millimolar concentrations of ornithine or micromolar concentrations of putrescine or by complementation with either glycosomal or cytosolic versions of ARG. Spermidine supplementation of the medium did not circumvent the polyamine auxotrophy of the Δarg line. Although ARG was found to be essential for ornithine and polyamine synthesis, ornithine decarboxylase appeared to be the rate-limiting enzyme for polyamine production. Mouse infectivity studies revealed that the Δarg lesion reduced parasite burdens in livers by an order of magnitude but had little impact on the numbers of parasites recovered from spleens. Thus, ARG is essential for proliferation of promastigotes but not intracellular amastigotes. Coupled with previous studies, these data support a model in which L. donovani amastigotes readily salvage ornithine and have some access to host spermidine pools, while host putrescine appears to be unavailable for salvage by the parasite.",
keywords = "Arginase, Leishmania, Polyamines",
author = "Boitz, {Jan M.} and Gilroy, {Caslin A.} and Olenyik, {Tamara D.} and Dustin Paradis and Jasmine Perdeh and Kristie Dearman and Davis, {Madison J.} and Phillip Yates and Yuexin Li and Michael Riscoe and Buddy Ullman and Roberts, {Sigrid C.}",
year = "2017",
doi = "10.1128/IAI.00554-16",
language = "English (US)",
volume = "85",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "1",

}

TY - JOUR

T1 - Arginase is essential for survival of Leishmania donovani promastigotes but not intracellular amastigotes

AU - Boitz, Jan M.

AU - Gilroy, Caslin A.

AU - Olenyik, Tamara D.

AU - Paradis, Dustin

AU - Perdeh, Jasmine

AU - Dearman, Kristie

AU - Davis, Madison J.

AU - Yates, Phillip

AU - Li, Yuexin

AU - Riscoe, Michael

AU - Ullman, Buddy

AU - Roberts, Sigrid C.

PY - 2017

Y1 - 2017

N2 - Studies of Leishmania donovani have shown that both ornithine decarboxylase and spermidine synthase, two enzymes of the polyamine biosynthetic pathway, are critical for promastigote proliferation and required for maximum infection in mice. However, the importance of arginase (ARG), the first enzyme of the polyamine pathway in Leishmania, has not been analyzed in L. donovani. To test ARG function in intact parasites, we generated Δarg null mutants in L. donovani and evaluated their ability to proliferate in vitro and trigger infections in mice. The Δarg knockout was incapable of growth in the absence of polyamine supplementation, but the auxotrophic phenotype could be bypassed by addition of either millimolar concentrations of ornithine or micromolar concentrations of putrescine or by complementation with either glycosomal or cytosolic versions of ARG. Spermidine supplementation of the medium did not circumvent the polyamine auxotrophy of the Δarg line. Although ARG was found to be essential for ornithine and polyamine synthesis, ornithine decarboxylase appeared to be the rate-limiting enzyme for polyamine production. Mouse infectivity studies revealed that the Δarg lesion reduced parasite burdens in livers by an order of magnitude but had little impact on the numbers of parasites recovered from spleens. Thus, ARG is essential for proliferation of promastigotes but not intracellular amastigotes. Coupled with previous studies, these data support a model in which L. donovani amastigotes readily salvage ornithine and have some access to host spermidine pools, while host putrescine appears to be unavailable for salvage by the parasite.

AB - Studies of Leishmania donovani have shown that both ornithine decarboxylase and spermidine synthase, two enzymes of the polyamine biosynthetic pathway, are critical for promastigote proliferation and required for maximum infection in mice. However, the importance of arginase (ARG), the first enzyme of the polyamine pathway in Leishmania, has not been analyzed in L. donovani. To test ARG function in intact parasites, we generated Δarg null mutants in L. donovani and evaluated their ability to proliferate in vitro and trigger infections in mice. The Δarg knockout was incapable of growth in the absence of polyamine supplementation, but the auxotrophic phenotype could be bypassed by addition of either millimolar concentrations of ornithine or micromolar concentrations of putrescine or by complementation with either glycosomal or cytosolic versions of ARG. Spermidine supplementation of the medium did not circumvent the polyamine auxotrophy of the Δarg line. Although ARG was found to be essential for ornithine and polyamine synthesis, ornithine decarboxylase appeared to be the rate-limiting enzyme for polyamine production. Mouse infectivity studies revealed that the Δarg lesion reduced parasite burdens in livers by an order of magnitude but had little impact on the numbers of parasites recovered from spleens. Thus, ARG is essential for proliferation of promastigotes but not intracellular amastigotes. Coupled with previous studies, these data support a model in which L. donovani amastigotes readily salvage ornithine and have some access to host spermidine pools, while host putrescine appears to be unavailable for salvage by the parasite.

KW - Arginase

KW - Leishmania

KW - Polyamines

UR - http://www.scopus.com/inward/record.url?scp=85009104912&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85009104912&partnerID=8YFLogxK

U2 - 10.1128/IAI.00554-16

DO - 10.1128/IAI.00554-16

M3 - Article

C2 - 27795357

AN - SCOPUS:85009104912

VL - 85

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 1

M1 - e00554-16

ER -