TY - JOUR
T1 - Leishmania donovani singly deficient in HGPRT, APRT or XPRT are viable in vitro and within mammalian macrophages
AU - Boitz, Jan M.
AU - Ullman, Buddy
N1 - Funding Information:
We thank Dr. Stephen Beverley of Washington University School of Medicine for generously providing the LdBob strain for these studies. We also thank Dr. Archie Bower, Dr. Michael Riscoe, and Dr. Jane Kelly for supplying the bone-derived macrophages and for their technical assistance. Finally, we thank Dr. Greg Matlashewski and Dr. Wen-Wei Zhang for providing us with the A2 antibody and technical assistance regarding its use. This work was supported in part by grants RO1 AI23682 from the National Institutes of Health (BU). JB has received financial support from the N.L. Tartar Research Fellowship from the Oregon Health and Science University.
PY - 2006/7
Y1 - 2006/7
N2 - Leishmania species express three phosphoribosyltransferase enzymes, hypoxanthine-guanine phosphoribosyltransferase (HGPRT), adenine phosphoribosyltransferase (APRT), and xanthine phosphoribosyltransferase (XPRT), which enable this genus to acquire purine nutrients from their hosts. To test whether any of these enzymes is essential for viability, transformation into amastigotes, and infectivity and proliferation within mammalian macrophages, Δhgprt, Δaprt, and Δxprt null mutants were created by targeted gene replacement within a virulent background of Leishmania donovani. Each of the three knockout strains was viable as promastigotes and axenic amastigotes and capable of maintaining an infection in bone marrow-derived murine macrophages. These data support the hypothesis that none of the three phosphoribosyltransferases is essential for purine salvage or viability by itself and that purine salvage occurs through multiple anabolic routes in both parasite life cycle stages. In addition these studies revealed the presence of an adenine aminohydrolase enzyme in L. donovani axenic amastigotes, an activity previously thought to be restricted to promastigotes.
AB - Leishmania species express three phosphoribosyltransferase enzymes, hypoxanthine-guanine phosphoribosyltransferase (HGPRT), adenine phosphoribosyltransferase (APRT), and xanthine phosphoribosyltransferase (XPRT), which enable this genus to acquire purine nutrients from their hosts. To test whether any of these enzymes is essential for viability, transformation into amastigotes, and infectivity and proliferation within mammalian macrophages, Δhgprt, Δaprt, and Δxprt null mutants were created by targeted gene replacement within a virulent background of Leishmania donovani. Each of the three knockout strains was viable as promastigotes and axenic amastigotes and capable of maintaining an infection in bone marrow-derived murine macrophages. These data support the hypothesis that none of the three phosphoribosyltransferases is essential for purine salvage or viability by itself and that purine salvage occurs through multiple anabolic routes in both parasite life cycle stages. In addition these studies revealed the presence of an adenine aminohydrolase enzyme in L. donovani axenic amastigotes, an activity previously thought to be restricted to promastigotes.
KW - Infectivity
KW - Metabolism
KW - Phosphoribosyltransferases
KW - Purines
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U2 - 10.1016/j.molbiopara.2006.02.015
DO - 10.1016/j.molbiopara.2006.02.015
M3 - Article
C2 - 16597468
AN - SCOPUS:33646926473
SN - 0166-6851
VL - 148
SP - 24
EP - 30
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
IS - 1
ER -