Depletion of UDP-Glucose and UDP-Galactose Using a Degron System Leads to Growth Cessation of Leishmania major

Interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal) by the UDP-Glc 4´-epimerase intimately connects the biosynthesis of these two nucleotide sugars. Their de novo biosynthesis involves transformation of glucose-6-phosphate into glucose-1-phosphate by the phosphoglucomutase and subse...

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Bibliographic Details
Published in:	PLoS neglected tropical diseases 2015-11, Vol.9 (11), p.e0004205
Main Authors:	Damerow, Sebastian, Hoppe, Carolin, Bandini, Giulia, Zarnovican, Patricia, Buettner, Falk F R, Buettner, Falk R, Lüder, Carsten G K, Ferguson, Michael A J, Routier, Françoise H
Format:	Article
Language:	eng
Subjects:	Apoptosis Biosynthesis Cell division Cellular control mechanisms Enzymes Gene Deletion Gene Expression Regulation Genetic aspects Glucose Growth Leishmania Leishmania major - growth & development Leishmania major - metabolism Ligands Metabolism Metabolites Parasites Parasitic diseases Properties Proteins Tropical diseases Uridine diphosphate Uridine Diphosphate Galactose - deficiency Uridine Diphosphate Galactose - metabolism Uridine Diphosphate Glucose - deficiency Uridine Diphosphate Glucose - metabolism UTP-Glucose-1-Phosphate Uridylyltransferase - genetics UTP-Hexose-1-Phosphate Uridylyltransferase - genetics
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Summary:	Interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal) by the UDP-Glc 4´-epimerase intimately connects the biosynthesis of these two nucleotide sugars. Their de novo biosynthesis involves transformation of glucose-6-phosphate into glucose-1-phosphate by the phosphoglucomutase and subsequent activation into UDP-Glc by the specific UDP-Glc pyrophosphorylase (UGP). Besides UGP, Leishmania parasites express an uncommon UDP-sugar pyrophosphorylase (USP) able to activate both galactose-1-phosphate and glucose-1-phosphate in vitro. Targeted gene deletion of UGP alone was previously shown to principally affect expression of lipophosphoglycan, resulting in a reduced virulence. Since our attempts to delete both UGP and USP failed, deletion of UGP was combined with conditional destabilisation of USP to control the biosynthesis of UDP-Glc and UDP-Gal. Stabilisation of the enzyme produced by a single USP allele was sufficient to maintain the steady-state pools of these two nucleotide sugars and preserve almost normal glycoinositolphospholipids galactosylation, but at the apparent expense of lipophosphoglycan biosynthesis. However, under destabilising conditions, the absence of both UGP and USP resulted in depletion of UDP-Glc and UDP-Gal and led to growth cessation and cell death, suggesting that either or both of these metabolites is/are essential.
ISSN:	1935-2735 1935-2727 1935-2735