title: Depletion of UDP-Glucose and UDP-Galactose Using a Degron System Leads to Growth Cessation of Leishmania major.
abstract: 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.
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Damerow
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Hoppe
authors.affiliation: Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Giulia
authors.initials: G
authors.lastname: Bandini
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Patricia
authors.initials: P
authors.lastname: Zarnovican
authors.forename: Falk F R
authors.initials: FF
authors.lastname: Buettner
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Falk R
authors.initials: FR
authors.lastname: Buettner
authors.affiliation: Institute for Medical Microbiology, Georg-August University, Goettingen, Germany.
authors.forename: Carsten G K
authors.initials: CG
authors.lastname: Lüder
authors.affiliation: Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee, United Kingdom.
authors.forename: Michael A J
authors.initials: MA
authors.lastname: Ferguson
authors.affiliation: Department of Cellular Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Françoise H
authors.initials: FH
authors.lastname: Routier
mesh_terms: D017353:Gene Deletion; D005786:Gene Expression Regulation; D018320:Leishmania major / Q000254:growth & development* / Q000378:metabolism*; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase / Q000235:genetics; D014531:Uridine Diphosphate Galactose / Q000172:deficiency* / Q000378:metabolism; D014532:Uridine Diphosphate Glucose / Q000172:deficiency* / Q000378:metabolism
chemical_list: D014531:Uridine Diphosphate Galactose; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D014532:Uridine Diphosphate Glucose