The role of phosphomannose isomerase in Leishmania mexicana glycoconjugate synthesis and virulence.
Phosphomannose isomerase (PMI) catalyzes the reversible interconversion of fructose 6-phosphate and mannose 6-phosphate, which is the first step in the biosynthesis of activated mannose donors required for the biosynthesis of various glycoconjugates. Leishmania species synthesize copious amounts of mannose-containing glycolipids and glycoproteins, which are involved in virulence of these parasitic protozoa. To investigate the role of PMI for parasite glycoconjugate synthesis, we have cloned the PMI gene (lmexpmi) from Leishmania mexicana, generated gene deletion mutants (Delta lmexpmi), and analyzed their phenotype. Delta lmexpmi mutants lack completely the high PMI activity found in wild type parasites, but are, in contrast to fungi, able to grow in media deficient for free mannose. The mutants are unable to synthesize phosphoglycan repeats [-6-Gal beta 1-4Man alpha 1-PO(4)-] and mannose-containing glycoinositolphospholipids, and the surface expression of the glycosylphosphatidylinositol-anchored dominant surface glycoprotein leishmanolysin is strongly decreased, unless the parasite growth medium is supplemented with mannose. The Delta lmexpmi mutant is attenuated in infections of macrophages in vitro and of mice, suggesting that PMI may be a target for anti-Leishmania drug development. L. mexicana Delta lmexpmi provides the first conditional mannose-controlled system for parasite glycoconjugate assembly with potential applications for the investigation of their biosynthesis, intracellular sorting, and function.
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Max-Planck-Institut für Biologie, Corrensstrasse 38, Tübingen 72076, Federal Republic of Germany.
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D000818:Animals; D001483:Base Sequence; D002855:Chromatography, Thin Layer; D006001:Glycoconjugates / Q000096:biosynthesis*; D007894:Leishmania mexicana / Q000378:metabolism* / Q000472:pathogenicity; D008264:Macrophages / Q000469:parasitology; D008358:Mannose / Q000378:metabolism / Q000494:pharmacology; D008359:Mannose-6-Phosphate Isomerase / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000096:biosynthesis; D017026:Swainsonine / Q000494:pharmacology; D014774:Virulence
D006001:Glycoconjugates; D011134:Polysaccharides; D008359:Mannose-6-Phosphate Isomerase; D008358:Mannose; D017026:Swainsonine