Biosynthesis and Anchoring of THY-1

Tartakoff, Alan M.

Abstract

Cancer cells may escape from immune surveillance if they mask or otherwise modulate the surface expression of critical surface antigens. The following proposal is an in-depth cell biological and biochemical study of the biosynthesis and modulation anchored to the lipid bilayer by a glycophospholipid (GPL) attached to the C- terminus of a truncated form of the primary translation product. A family of Thy-1 negative lymphoma mutants produced by negative immunoselection has been show that representatives of groups A-C secrete anchor-deficient, hydrophilic forms of Thy-1 (see enclosed reprint and preprint). The proposed studies aim: I. To use wild-type cells to establish the subcellular site(s) of anchor addition, to set up a microsomal for in vitro anchor addition, and to learn how many lymphoma proteins bear a GPL. II. To use the mutant lymphomas for investigation of anchor biosynthetic lesions and for comparison with cells recovered from paroxysmal nocturnal hemoglobinuria (PNH) patients. III. To use the approach to transfection to anchor both Thy-1 and secretory proteins to the surface of CHO cells via a GPL anchor. Taken together, these studies will elucidate normal GPL anchor biosynthesis and evaluate the hypothesis that secretion of Thy-1 by the mutant cells is the result of lesions in the pathway of anchor biosynthesis.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
• Type: Research Program Projects (P01)
• Project #: 5P01DK038181-02
• Application #: 3918044
• Support Year: 2
• Fiscal Year: 1988

Institution

• Name: Case Western Reserve University
• DUNS #: 077758407
• City: Cleveland
• State: OH
• Country: United States
• Zip Code: 44106

Publications

Chen, R; Knez, J J; Merrick, W C et al. (2001) Comparative efficiencies of C-terminal signals of native glycophosphatidylinositol (GPI)-anchored proproteins in conferring GPI-anchoring. J Cell Biochem 84:68-83

Wongkajornsilp, A; Sevlever, D; Rosenberry, T L (2001) Metabolism of exogenous sn-1-alkyl-sn-2-lyso-glucosaminyl-phosphatidylinositol in HeLa D cells: accumulation of glucosaminyl(acyl)phosphatidylinositol in a metabolically inert compartment. Biochem J 359:305-13

Premkumar, D R; Fukuoka, Y; Sevlever, D et al. (2001) Properties of exogenously added GPI-anchored proteins following their incorporation into cells. J Cell Biochem 82:234-45

Sevlever, D; Pickett, S; Mann, K J et al. (1999) Glycosylphosphatidylinositol-anchor intermediates associate with triton-insoluble membranes in subcellular compartments that include the endoplasmic reticulum. Biochem J 343 Pt 3:627-35

Chen, A; Meyerson, H J; Salvekar, A et al. (1998) Non-glycosylated human B7-1(CD80) retains the capacity to bind its counter-receptors. FEBS Lett 428:127-34

Chen, R; Walter, E I; Parker, G et al. (1998) Mammalian glycophosphatidylinositol anchor transfer to proteins and posttransfer deacylation. Proc Natl Acad Sci U S A 95:9512-7

Kraus, D; Medof, M E; Mold, C (1998) Complementary recognition of alternative pathway activators by decay-accelerating factor and factor H. Infect Immun 66:399-405

Sevlever, D; Schiemann, D; Guidubaldi, J et al. (1997) Accumulation of glucosaminyl(acyl)phosphatidylinositol in an S3 HeLa subline expressing normal dolicholphosphomannose synthase activity. Biochem J 321 ( Pt 3):837-44

Yu, J; Nagarajan, S; Knez, J J et al. (1997) The affected gene underlying the class K glycosylphosphatidylinositol (GPI) surface protein defect codes for the GPI transamidase. Proc Natl Acad Sci U S A 94:12580-5

Ramalingam, S; Maxwell, S E; Medof, M E et al. (1996) COOH-terminal processing of nascent polypeptides by the glycosylphosphatidylinositol transamidase in the presence of hydrazine is governed by the same parameters as glycosylphosphatidylinositol addition. Proc Natl Acad Sci U S A 93:7528-33