Adaptive responses to purine starvation in Leishmania donovani

Nicola Carter, Phillip Yates, Sarah K. Gessford, Sean R. Galagan, Scott Landfear, Buddy Ullman

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Starvation of Leishmania donovani parasites for purines leads to a rapid amplification in purine nucleobase and nucleoside transport. Studies with nucleoside transport-deficient L. donovani indicate that this phenomenon is mediated by the nucleoside transporters LdNT1 and LdNT2, as well as by the purine nucleobase transporter LdNT3. The escalation in nucleoside transport cannot be ascribed to an increase in either LdNT1 or LdNT2 mRNA. However, Western analyses on parasites expressing epitope-tagged LdNT2 revealed a marked upregulation in transporter protein at the cell surface. Kinetic investigations of LdNT1 and LdNT2 activities from purine-replete and purine-starved cells indicated that both transporters exhibited significant increases in V max for their ligands under conditions of purine-depletion, although neither transporter displayed an altered affinity for its respective ligands. Concomitant with the increase in purine nucleoside and nucleobase transport, the purine salvage enzymes HGPRT, XPRT and APRT were also upregulated, suggesting that under conditions where purines are limiting, Leishmania parasites remodel their purine metabolic pathway to maximize salvage. Moreover, qRT-PCR analyses coupled with cycloheximide inhibition studies suggest that the underlying molecular mechanism for this augmentation in purine salvage occurs post-transcriptionally and is reliant on de novo protein synthesis.

Original languageEnglish (US)
Pages (from-to)92-107
Number of pages16
JournalMolecular Microbiology
Volume78
Issue number1
DOIs
StatePublished - Oct 2010

Fingerprint

Leishmania donovani
Starvation
Purine Nucleosides
Purines
Parasites
Nucleosides
Nucleoside Transport Proteins
Ligands
Hypoxanthine Phosphoribosyltransferase
Leishmania
Cycloheximide
Metabolic Networks and Pathways
purine
Epitopes
Membrane Proteins
Up-Regulation
Polymerase Chain Reaction
Messenger RNA
Enzymes

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Adaptive responses to purine starvation in Leishmania donovani. / Carter, Nicola; Yates, Phillip; Gessford, Sarah K.; Galagan, Sean R.; Landfear, Scott; Ullman, Buddy.

In: Molecular Microbiology, Vol. 78, No. 1, 10.2010, p. 92-107.

Research output: Contribution to journalArticle

Carter, Nicola ; Yates, Phillip ; Gessford, Sarah K. ; Galagan, Sean R. ; Landfear, Scott ; Ullman, Buddy. / Adaptive responses to purine starvation in Leishmania donovani. In: Molecular Microbiology. 2010 ; Vol. 78, No. 1. pp. 92-107.
@article{3388500120c9438690e4d15cff1dd822,
title = "Adaptive responses to purine starvation in Leishmania donovani",
abstract = "Starvation of Leishmania donovani parasites for purines leads to a rapid amplification in purine nucleobase and nucleoside transport. Studies with nucleoside transport-deficient L. donovani indicate that this phenomenon is mediated by the nucleoside transporters LdNT1 and LdNT2, as well as by the purine nucleobase transporter LdNT3. The escalation in nucleoside transport cannot be ascribed to an increase in either LdNT1 or LdNT2 mRNA. However, Western analyses on parasites expressing epitope-tagged LdNT2 revealed a marked upregulation in transporter protein at the cell surface. Kinetic investigations of LdNT1 and LdNT2 activities from purine-replete and purine-starved cells indicated that both transporters exhibited significant increases in V max for their ligands under conditions of purine-depletion, although neither transporter displayed an altered affinity for its respective ligands. Concomitant with the increase in purine nucleoside and nucleobase transport, the purine salvage enzymes HGPRT, XPRT and APRT were also upregulated, suggesting that under conditions where purines are limiting, Leishmania parasites remodel their purine metabolic pathway to maximize salvage. Moreover, qRT-PCR analyses coupled with cycloheximide inhibition studies suggest that the underlying molecular mechanism for this augmentation in purine salvage occurs post-transcriptionally and is reliant on de novo protein synthesis.",
author = "Nicola Carter and Phillip Yates and Gessford, {Sarah K.} and Galagan, {Sean R.} and Scott Landfear and Buddy Ullman",
year = "2010",
month = "10",
doi = "10.1111/j.1365-2958.2010.07327.x",
language = "English (US)",
volume = "78",
pages = "92--107",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Adaptive responses to purine starvation in Leishmania donovani

AU - Carter, Nicola

AU - Yates, Phillip

AU - Gessford, Sarah K.

AU - Galagan, Sean R.

AU - Landfear, Scott

AU - Ullman, Buddy

PY - 2010/10

Y1 - 2010/10

N2 - Starvation of Leishmania donovani parasites for purines leads to a rapid amplification in purine nucleobase and nucleoside transport. Studies with nucleoside transport-deficient L. donovani indicate that this phenomenon is mediated by the nucleoside transporters LdNT1 and LdNT2, as well as by the purine nucleobase transporter LdNT3. The escalation in nucleoside transport cannot be ascribed to an increase in either LdNT1 or LdNT2 mRNA. However, Western analyses on parasites expressing epitope-tagged LdNT2 revealed a marked upregulation in transporter protein at the cell surface. Kinetic investigations of LdNT1 and LdNT2 activities from purine-replete and purine-starved cells indicated that both transporters exhibited significant increases in V max for their ligands under conditions of purine-depletion, although neither transporter displayed an altered affinity for its respective ligands. Concomitant with the increase in purine nucleoside and nucleobase transport, the purine salvage enzymes HGPRT, XPRT and APRT were also upregulated, suggesting that under conditions where purines are limiting, Leishmania parasites remodel their purine metabolic pathway to maximize salvage. Moreover, qRT-PCR analyses coupled with cycloheximide inhibition studies suggest that the underlying molecular mechanism for this augmentation in purine salvage occurs post-transcriptionally and is reliant on de novo protein synthesis.

AB - Starvation of Leishmania donovani parasites for purines leads to a rapid amplification in purine nucleobase and nucleoside transport. Studies with nucleoside transport-deficient L. donovani indicate that this phenomenon is mediated by the nucleoside transporters LdNT1 and LdNT2, as well as by the purine nucleobase transporter LdNT3. The escalation in nucleoside transport cannot be ascribed to an increase in either LdNT1 or LdNT2 mRNA. However, Western analyses on parasites expressing epitope-tagged LdNT2 revealed a marked upregulation in transporter protein at the cell surface. Kinetic investigations of LdNT1 and LdNT2 activities from purine-replete and purine-starved cells indicated that both transporters exhibited significant increases in V max for their ligands under conditions of purine-depletion, although neither transporter displayed an altered affinity for its respective ligands. Concomitant with the increase in purine nucleoside and nucleobase transport, the purine salvage enzymes HGPRT, XPRT and APRT were also upregulated, suggesting that under conditions where purines are limiting, Leishmania parasites remodel their purine metabolic pathway to maximize salvage. Moreover, qRT-PCR analyses coupled with cycloheximide inhibition studies suggest that the underlying molecular mechanism for this augmentation in purine salvage occurs post-transcriptionally and is reliant on de novo protein synthesis.

UR - http://www.scopus.com/inward/record.url?scp=77957235442&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957235442&partnerID=8YFLogxK

U2 - 10.1111/j.1365-2958.2010.07327.x

DO - 10.1111/j.1365-2958.2010.07327.x

M3 - Article

C2 - 20923417

AN - SCOPUS:77957235442

VL - 78

SP - 92

EP - 107

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 1

ER -