The primary glycosylation defect in class E Thy-1-negative mutant mouse lymphoma cells is an inability to synthesize dolichol-P-mannose.

Chapman, Ann E.; Fujimoto, K; Kornfeld, Stuart

doi:10.1016/s0021-9258(19)85510-2

Cited by 234 publications

(19 citation statements)

References 23 publications

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“…In addition, four of five types of mutant lymphoma cells isolated because of decreased expression of Thy-1 antigen show altered glycosylation of this molecule (Trowbridge et al, 1978). One of these classes of mutant, Thy-1 E, has a defect in dolichol-mannose-phosphate synthetase activity that is very similar to that in the mannose-6-phosphate receptor defective CHO mutant (Chapman et al, 1980). The close associations between glycosylation defects and altered expression of the LDL receptor, the mannose-6-phosphate receptor, and the Thy-1 antigen emphasize the importance of carbohydrate chains for the expression and function of some cell surface glycoproteins.…”

Section: Discussionmentioning

confidence: 99%

Three types of low density lipoprotein receptor-deficient mutant have pleiotropic defects in the synthesis of N-linked, O-linked, and lipid-linked carbohydrate chains.

Kingsley¹,

Kozarsky²,

Segal³

et al. 1986

The Journal of Cell Biology

178

236

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Abstract. Biochemical, immunological, and genetic techniques were used to investigate the genetic defects in three types of low density lipoprotein (LDL) receptor-deficient hamster cells. The previously isolated ldlB, ldlC, and ldlD mutants all synthesized essentially normal amounts of a 125,000-D precursor form of the LDL receptor, but were unable to process this receptor to the mature form of 155,000 D. Instead, these mutants produced abnormally small, heterogeneous receptors that reached the cell surface but were rapidly degraded thereafter. The abnormal sizes of the LDL receptors in these cells were due to defective processing of the LDL receptor's N-and O-linked carbohydrate chains. Processing defects in these cells appeared to be general since the ldlB, ldlC, and ldlD mutants also showed defective glycosylation of a viral glycoprotein, alterations in glycolipid synthesis, and changes in resistance to several toxic lectins. Preliminary structural studies suggested that these cells had defects in multiple stages of the Golgi-associated processing reactions responsible for synthesis of glycolipids and in the N-linked and O-linked carbohydrate chains of glycoproteins. Comparisons between the ldl mutants and a large number of previously isolated CHO glycosylation defective mutants showed that the genetic defects in ldlB, ldlC, and ldlD cells were unique and that only very specific types of carbohydrate alteration could dramatically affect LDL receptor function.

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Section: Discussionmentioning

confidence: 99%

Three types of low density lipoprotein receptor-deficient mutant have pleiotropic defects in the synthesis of N-linked, O-linked, and lipid-linked carbohydrate chains.

Kingsley¹,

Kozarsky²,

Segal³

et al. 1986

The Journal of Cell Biology

178

236

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“…BW5147(Thy-1 -a) and BW5147(Thy-1 -e) are Thy-1 -mutant cell lines derived from BW5147(Thy-1+) wild type (34,35) . Both class A and E mutants are defective in the anchoring of membrane Thy-1 glycoproteins via the phosphatidylinositol-containing glycolipid anchor ofThy-1 (36), and the class E mutant has a mutation in the gene coding for GDP-mannose :dolichol-P-mannosyltransferase, which presumably affects the biosynthesis of glycolipid tails in addition to the biosynthesis ofWinked high-mannose oligosaccharides (36,37) . The AKR1(Thy-1 -d) cell line is a class D mutant from AKR1 (Thy-1+) wild type.…”

Section: Methodsmentioning

confidence: 99%

“…To more directly study the relationship of the determinants defined by the ATA antibodies ofgroups I and II to the Thy-1 molecule, the staining of these antibodies on a panel of cell lines was examined. BW5147 is a Thy-1+ lymphoma derived from AKR/J (Thy-1 .1) mice, while BW5147 (Thy-1 -a) and BW5147 (Thy-1-e) are Thy-1 -mutants with defects in genes affecting post-translational steps necessary for cell surface expression of the Thy-1 glycoprotein (and other cell surface molecules anchored in the cell membrane through a glycophospholipid moiety) (34)(35)(36)(37) . AKRl is also a Thy-1+ lymphoma derived from AKR/J mice, while AKRl (Thy-1 -d) is a Thy-1 -mutant that has undergone deletion of a portion of the Thy-1 structural gene (38).…”

mentioning

confidence: 99%

Natural autoantibodies to thymocytes: origin, VH genes, fine specificities, and the role of Thy-1 glycoprotein.

Hayakawa

Carmack

Hyman

et al. 1990

The Journal of Experimental Medicine

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Summary15 SM/J mouse hybridoma antibodies that show antithymocyte autoantibody (ATA) activity by immunofluorescence staining were studied. Half of these antibodies react with determinants whose expression is associated with Thy-1, as shown by blocking experiments with antiThy-1 and loss of reactivity with Thy-1 -mutant cell lines . The Thy-1 dependence of three of these ATA is further confirmed by their reexpression on a Thy-1 gene transfectant . However, the remaining antibodies exhibited binding that showed little or no dependence on Thy-1. Furthermore, we find that most ATA derives from the Ly-1 B subpopulation, as demonstrated by lipopolisaccharideinduced ATA secretion in vitro and by comparison of ATA hybridoma frequencies. V region gene sequence data of 14 monoclonal ATA from Ly-1 B cell-derived hybridomas reveal the utilization of nine V genes belonging to four different V H families (J558, 3609, Q52, and Vgam3.8) . While we find that two of these hybridomas arose from a clonal expansion, we also find four examples of a 3609 family V gene utilized in clonally independent lines showing similar specificity. Yet another example of identical V gene usage by clonally unrelated cells is found in two J558 ATA of a distinct fine specificity. These data suggest that the enrichment of ATA B cells in the Ly-1 B subset is primarily due to repeated independent recruitment of B cells by antigen resulting in the expression of a restricted set of VH genes.

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“…4). The basic tenet of both models is that GPI-anchored proteins are committed to the secretory pathway by default, and are retained in the plasma membrane only by GPI addition (11,13,21,67). In the first, the GPI-addition-cleavage model, gp63 was presumed to initially acquire a GPI anchor in the ER.…”

Section: Gpi-plc Causes Rapid Secretion Of Gpi-anchorless Gp63: Gpi Biosynthesis Is Initiated On the Cytoplasmic Side Of The Endoplasmic mentioning

confidence: 99%

A glycosylphosphatidylinositol (GPI)-negative phenotype produced in Leishmania major by GPI phospholipase C from Trypanosoma brucei: topography of two GPI pathways

Mensa-Wilmot

LeBowitz

Chang

et al. 1994

The Journal of Cell Biology

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Abstract. The major surface macromolecules of the protozoan parasite Leishmania major, gp63 (a metalloprotease), and lipophosphoglycan (a polysaccharide), are glycosylphosphatidylinositol (GPI) anchored. We expressed a cytoplasmic glycosylphosphatidylinositol phospholipase C (GPI-PLC) in L. major in order to examine the topography of the protein-GPI and polysaccharide-GPI pathways. In L. major cells expressing GPI-PLC, cell-associated gp63 could not be detected in immunoblots. Pulse-chase analysis revealed that gp63 was secreted into the culture medium with a half-time of 5.5 h. Secreted gp63 lacked anti-cross reacting determinant epitopes, and was not metabolically labeled with [3H]ethanolamine, indicating that it never received a GPI anchor. Further, the quantity of putative protein-GPI intermediates decreased ,,ol0-fold. In striking contrast, lipophosphoglycan levels were unaltered. However, GPI-PLC cleaved polysaccharide-GPI intermediates (glycoinositol phospholipids) in vitro. Thus, reactions specific to the polysaccharide-GPI pathway are comparianentalized in vivo within the endoplasmic reticulum, thereby sequestering polysaccharide-GPI intermediates from GPI-PLC cleavage. On the contrary, protein-GPI synthesis at least up to production of Man(lat)Man(lot4)GlcN-(lat)-myo-inositol-l-phospholipid is cytosolic. To our knowledge this represents the first use of a catabolic enzyme in vivo to elucidate the topography of biosynthetic pathways.GPI-PLC causes a protein-GPI-negative phenotype in L. major, even when genes for GPI biosynthesis are functional. This phenotype is remarkably similar to that of some GPI mutants of mammalian cells: implications for paroxysmal nocturnal hemoglobinuria and Thy-l-negative T-lymphoma are discussed. LYCOSYLPHOSPHATIDYLINOSITOL (GP1) t anchors attach a diverse group of macromolecules to membranes in eukaryotes (see 19, 22 for recent reviews). The plasma membrane of the protozoan parasite Trypmwsoma brucei is covered with about 107 molecules of the variant surface glycoprotein (VSG), a GPI-anchored molePlease address all correspondence to Dr. K. Mensa-Wilmot, Department of Zoology, University of Georgia, Athens, Georgia 30602.1. Abbreviations used in this paper: CRD, cross-reacting determinant; dol-P-Man, dolichol-phosphoryl-mannose; eGPI-PLC, recombinant glycosylphosphatidylinositol phospholipase C; EtN, ethanolamine; gp63, 63-kD GPI-anchored glycoprotein of Leishmania parasites; GIPL, glycoinositol phospholipid; GPI, glycosylphosphatidylinositol; GPI-PLC, glycosylphosphatidylinositol phospholipnse C; Gall, Galactofuranose; GleN, glucosamine; GIcNAC, N-acetyl glucosamine; LP-1, putative protein-GPI precursor (possibly EtN-phospho-ManrGlcN-PI); LP-2, putative protein-GPl precursor; LPG, lipophosphoglycan; Man, mannose; mfVSG, membraneform VSG; PI, phosphatidylinositol; PNH, paroxysmal nocturnal hemoglobinuria; UDP-GIcNAc, uridine 5'-diphospho N-acetylglucosamine; VSG, variant surface glycoprotein of Trypanosoma brucei.cule. The VSG GPI consists of dimyristoylphosphatidylinositol l...

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The primary glycosylation defect in class E Thy-1-negative mutant mouse lymphoma cells is an inability to synthesize dolichol-P-mannose.

Cited by 234 publications

References 23 publications

Three types of low density lipoprotein receptor-deficient mutant have pleiotropic defects in the synthesis of N-linked, O-linked, and lipid-linked carbohydrate chains.

Three types of low density lipoprotein receptor-deficient mutant have pleiotropic defects in the synthesis of N-linked, O-linked, and lipid-linked carbohydrate chains.

Natural autoantibodies to thymocytes: origin, VH genes, fine specificities, and the role of Thy-1 glycoprotein.

A glycosylphosphatidylinositol (GPI)-negative phenotype produced in Leishmania major by GPI phospholipase C from Trypanosoma brucei: topography of two GPI pathways

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