Abstract
Familial hypercholesterolemia (FH) is an autosomal dominant disease caused by mutations in the low density lipoprotein (LDL) receptor gene. Here, we characterize an LDL receptor mutation that is associated with a distinct haplotype and that causes FH in the Jewish Sephardic population originating from Safed, a town in northern Israel. The mutation was found in eight FH families originating from this community comprising 10% of heterozygote FH index cases screened in Israel. The mutation was not found in four additional FH heterozygotes whose hypercholesterolemia co-segregated with an identical LDL receptor gene haplotype. A guanine to cytosine substitution results in a missense mutation (asp147 to his) in the fourth repeat of the binding domain encoded by exon 4 of the LDL receptor gene. The mutant receptor protein was synthesized in cultured cells as a 120kDa precursor form that failed to undergo normal processing to a mature cell surface form. Most of the receptor precursors were degraded in the endoplasmic reticulum. The small number of mutant receptors on the cell surface were unable to bind LDL or β very low density lipoprotein. The abnormal behavior of the mutant receptor was reproduced by site-directed mutagenesis and expression of the mutant protein in CHO cells. The mutation can be diagnosed by allele-specific oligonucleotide hybridization of polymerase chain reaction amplified DNA from FH patients.
Similar content being viewed by others
References
Aalto-Setälä K, Helve E, Kovanen PT, Kontula K (1989) Finnish type of low density lipoprotein receptor gene mutation (FH-Helsinki) deletes exons encoding the carboxyl-terminal part of the receptor and creates an internalization-defective phenotype. J Clin Invest 84:499–505
Aulinskas TH, Westhuyzen DR van der, Coetzee GA (1983) Ascorbate increases the number of the low density lipoprotein receptors in cultured arterial smooth muscle cells. Atherosclerosis 47:159–171
Beisiegel U, Schneider WJ, Goldstein JL, Anderson RGW, Brown MS (1981) Monoclonal antibodies to the low density lipoprotein receptor as probes for the study of receptor-mediated endocytosis and the genetics of familial hypercholesterolemia. J Biol Chem 256:11923–11931
Berkman N, Weir BS, Pressman Schwartz S, Reshef A, Leitersdorf E (1992) Haplotype analysis at the low density lipoprotein receptor locus: application for the analysis of familial hypercholesterolemia in Israel. Hum Genet 88:405–410
Brown MS, Goldstein JL (1986) A receptor-mediated pathway for cholesterol homeostasis. Science 232:34–47
Chirgwin MJ, Przybyla EA, McDonald JR, Rutter JW (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299
Cooper DN, Krawczak M (1990) The mutationul spectrum of single base-pair substitutions causing human genetic diseases: patterns and predictions. Hum Genet 85:55–74
Daniel TO, Schneider WJ, Goldstein JL, Brown MS (1983) Visualization of lipoprotein receptors by blotting. J Biol Chem 258:4606–4611
Davis CG, Elhammer A, Russell DW, Schneider WJ, Kornfeld S, Brown MS, Goldstein JL (1986) Deletion of clustered O-linked carbohydrates does not impair function of low density lipoprotein receptor in transfected fibroblasts. J Biol Chem 261:2828–2838
Driel IR van, Davis CG, Glodstein JL, Brown MS (1987) Self-association of the low density lipoprotein receptor mediated by the cytoplasmic domain. J Biol Chem 262:16127–16134
Friedewald WT, Levy RI, Fredrickson DS (1972) Estimation of the concentration of low density lipoprotein cholesterol in plasma without use of the preparative ultracentrifuge. Clin Chem 18:499–502
Gething M-J, Sambrook J (1992) Protein folding in the cell. Nature 355:33–45
Goldstein JL, Brown MS (1989) Familial hypercholesterolemia. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic basis of inherited disease. 6th edn. McGraw-Hill, New York, pp 1215–1250
Goldstein JL, Basu SK, Brown MS (1983) Receptor-mediated endocytosis of low density lipoprotein in cultured cells. Methods Enzymol 98:241–260
Grant KI, Casciola LAF, Coetzee GA, Sanan DA, Gcvers W, Westhuyzen DR van der (1990) Ammonium chloride causes reversible inhibition of low density lipoprotein receptor recycling and accelerates receptor degradation. J Biol Chem 256:4041–4047
Hobbs HH, Brown MS, Goldstein JL, Russell DW (1986) Deletion of exon encoding cysteine-rich repeat of the LDL receptor alters its binding specificity in a subject with familial hypercholesterolemia. J Biol Chem 261:13114–13120
Hobbs HH, Brown MS, Russell DW, Davignon J, Goldstein JL (1987) Deletion in the gene for the low-density lipoprotein receptor in a majority of French Canadians with familial hypercholesterolemia. N Engl J Med 317:734–737
Hobbs HH, Russell DW, Brown MS, Goldstein JL (1990) The LDL receptor locus in familial hypercholesterolemia: mutationl analysis of a membrane protein. Annu Rev Genet 24:133–170
Kingsley DM, Krieger M (1984) Receptor mediated endocytosis of low density lipoprotein: somatic cell mutants define multiple genes required for expression of surface-receptor activity. Proc Natl Acad Sci USA 81:5454–5458
Klausner RD, Sitia R (1990) Protein degradation in the endoplasmic reticulum. Cell 62:611–614
Kunkel TA (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492
Landsberger D, Meiner V, Reshef A, Levy Y, Westhuyzen DR van der, Coetzee GA, Leitersdorf E (1992) A nonsense mutation in the LDL receptor gene leads to familial hypercholesterolemia in the Druze sect. Am J Hum Genet 50:427–433
Lehrman MA, Schneider WJ, Brown MS, Davis CG, Elhammer A, Russell DW, Goldstein JL (1987) The Lebanese allele at the LDL receptor gene locus: nonsense mutation produces truncated receptor that is retained in the endoplasmic reticulum. J Biol Chem 262:401–410
Leitersdorf E, Chakravarti A, Hobbs HH (1989a) Polymorphic DNA haplotypes at the LDL receptor locus. Am J Hum Genet 44:409–421
Leitersdorf E, Westhuyzen DR van der, Coetzee Ga, Hobbs HH (1989b) Two common low density lipoprotein gene mutations cause familial hypercholesterolemia in Afrikaners. J Clin Invest 84:954–961
Leitersdorf E, Tobin EJ, Davignon J, Hobbs HH (1990) Common low-density lipoprotein receptor mutations in the French Canadian population. J Clin Invest 85:1014–1023
Leitersdorf E, Friedlander Y, Bard JM, Fruchart JC, Eisenberg S, Stein Y (1991) Diverse effect of ethnicity on plasma lipoprotein(a) levels in heterozygote patients with familial hypercholesterolemia. J Lipid Res 32:1513–1519
Meiner V, Landsberger D, Berkman N, Reshef A, Segal P, Seftel HC, Westhuyzen DR van der, Jeenah MS, Coetzee GA, Leitersdorf E (1991) A common Lithuanian mutation causing familial hypercholesterolemia in Ashkenazi Jews. Am J Hum Genet 49:443–449
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78
Montefiore M (ed) (1839) A census of the Jews of Eretz Israel. The Dinur Center, The Hebrew University, Jerusalem, Israel
Oppenheim A, Friedlander Y, Dann EJ, Berkman N, Pressman Schwartz S, Leitersdorf E (1991) Hypercholesterolemia in five Israeli Christian-Arab kindreds is caused by the “Lebanese” allele at the LDL receptor gene locus as well as by an additional independent major factor. Hum Genet 88:75–84
Orci L, Tagoyo M, Amherdt M, Perrelet A, Donaldson Jg, Lippincott-Schwartz J, Klausner RD, Rothman JE (1991) Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrin coated buds on Golgi cisternae. Cell 64:1183–1195
Russell DW, Brown MS, Goldstein JL (1989) Different combinations of cysteine-rich repeats mediate binding of low density lipoprotein receptor to two different proteins. J Biol Chem 264:21682–21688
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuuchi RH, Horn GT, Mullis KB, Erlich HA (1988) Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Tolleshaug H, Goldstein JL, Schneider WJ, Brown MS (1982) Posttranslational processing of the LDL receptor and its genetic disruption in familial hypercholesterolemia. Cell 30:715–724
Utermann G (1989) The mysteries of lipoprotein(a). Science 246:904–910
Yamamoto T, Davis CG, Brown MS, Schneider WJ, Casey ML, Goldstein JL, Russell DW (1984) The human LDL receptor: a cysteine-rich protein with multiple alu sequences in its mRNA. Cell 39:27–38
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Leitersdorf, E., Reshef, A., Meiner, V. et al. A missense mutation in the low density lipoprotein receptor gene causes familial hypercholesterolemia in Sephardic Jews. Hum Genet 91, 141–147 (1993). https://doi.org/10.1007/BF00222714
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00222714