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Mannosidase 2, alpha 1 deficiency is associated with ricin resistance in embryonic stem (ES) cells

Wang, Wei and Hale, Christine and Goulding, Dave and Haslam, Stuart and Tissot, Berangere and Lindsay, Christopher and Michell, Stephen and Titball, Rick and Yu, Jun and Toribio, Ana Luisa and Rossi, Raffaella and Dell, Anne and Bradley, Allan and Dougan, Gordon (2011) Mannosidase 2, alpha 1 deficiency is associated with ricin resistance in embryonic stem (ES) cells. PLoS One, 6 (8). ISSN 1932-6203

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Abstract

Host gene products required for mediating the action of toxins are potential targets for reversing or controlling their pathogenic impact following exposure. To identify such targets libraries of insertional gene-trap mutations generated with a PiggyBac transposon in Blm-deficient embryonic stem cells were exposed to the plant toxin, ricin. Resistant clones were isolated and genetically characterised and one was found to be a homozygous mutant of the mannosidase 2, alpha 1 (Man2α1) locus with a matching defect in the homologous allele. The causality of the molecular lesion was confirmed by removal of the transposon following expression of PB-transposase. Comparative glycomic and lectin binding analysis of the Man2α1 (-/-) ricin resistant cells revealed an increase in the levels of hybrid glycan structures and a reduction in terminal β-galactose moieties, potential target receptors for ricin. Furthermore, naïve ES cells treated with inhibitors of the N-linked glycosylation pathway at the mannosidase 2, alpha 1 step exhibited either full or partial resistance to ricin. Therefore, we conclusively identified mannosidase 2, alpha 1 deficiency to be associated with ricin resistance.