Deciphering the Glycosylome of Dystroglycanopathies Using Haploid Screens for Lassa Virus Entry

Abstract

Viruses and Congenital Disorders Mutations in genes involved in α-dystroglycan O-linked glycosylation result in posttranslation modifications associated with the congenital disease Walker-Warburg syndrome (WWS). This cellular modification is also required for efficient Lassa virus infection of cells. Jae et al. (p. 479 , published online 21 March) screened for genes involved in O-glycosylation that affected Lassa virus infection and identified candidates involved in glycosylation. Individuals from different pedigrees exhibiting WWS had unique mutations among genes identified in the genetic screen. Thus, comprehensive forward genetic screens can be used to define the genetic architecture of a complex disease.

Enterovirus D68 receptor requirements unveiled by haploid genetics

“…ST3GAL4 KO and SLC35A1 KO HAP1 cells have been described (31). CMAS KO cells were obtained from Haplogen GmbH.…”

“…After selection, surviving cells were expanded and used for genomic DNA isolation. Insertion sites identified in cells selected with EV-D68 (yielding 414,290 unique gene-trap insertions mapped to genes) and a population of matched control cells of comparable complexity (495,679 unique gene-trap insertions mapped to genes) were aligned to the human genome not filtering for close reads (31). Subsequently, disruptive insertion sites (in sense orientation of the affected gene or mapping to exons) in significantly identified genes were compared in the two cell populations, and P values for enrichment were calculated using a Fisher's exact test as described previously (31).…”

“…Insertion sites identified in cells selected with EV-D68 (yielding 414,290 unique gene-trap insertions mapped to genes) and a population of matched control cells of comparable complexity (495,679 unique gene-trap insertions mapped to genes) were aligned to the human genome not filtering for close reads (31). Subsequently, disruptive insertion sites (in sense orientation of the affected gene or mapping to exons) in significantly identified genes were compared in the two cell populations, and P values for enrichment were calculated using a Fisher's exact test as described previously (31). Disruptive insertion sites in virus-selected and control cells were plotted onto the RefSeq gene bodies for the following transcripts: NM_001497 (B4GALT1), NM_018686 (CMAS), NM_001128227 (GNE), NM_002410 (MGAT5), NM_018946 (NANS), NM_006416 (SLC35A1), NM_005660 (SLC35A2), NM_006278 (ST3GAL4), and NM_173216.2 (ST6GAL1).…”

“…HAP1 cells were gene-trap mutagenized as described previously (31). Following expansion, 10 8 mutagenized cells were exposed to EV-D68 Fermon (moi 3).…”

Mining the O-mannose glycoproteome reveals cadherins as major O-mannosylated glycoproteins

“…found on Ser and Thr residues was initially thought to be restricted to yeast and fungi, and only within the last two decades has it been shown that this glycosylation occurs in metazoans (1,2). O-Man glycosylation of the basement membrane glycoprotein α-dystroglycan (α-DG) is essential for assembly and function of the dystrophin-glycoprotein complex that links the cytoskeleton with the extracellular matrix, and deficiencies in all of the enzymes involved in the O-Man glycosylation underlie congenital muscular dystrophies (dystroglycanopathies) (3)(4)(5).…”