Human Genetics

, Volume 92, Issue 1, pp 6–10 | Cite as

Screening for point mutations in exon 10 of the low density lipoprotein receptor gene by analysis of single-strand conformation polymorphisms: detection of a nonsense mutation — FH469→Stop

  • Trond P. Leren
  • Kari Solberg
  • Olaug K. Rødningen
  • Oddveig Røsby
  • Serena Tonstad
  • Leiv Ose
  • Kåre Berg
Original Investigations
Received:
Revised:

Abstract

DNA from 40 unrelated familial hypercholesterolemia (FH) heterozygotes were subjected to analyses of single-strand conformation polymorphisms (SSCPs) of exon 10 of the low density lipoprotein receptor (LDLR) gene. Four different SSCP patterns were observed. The underlying mutations were characterized by DNA sequencing. Three of the patterns represented the three genotypes of a recently described sense mutation in codon 450. A method based upon the polymerase chain reaction (PCR) was developed to analyze this mutation. The frequencies of the wild-type (G at nucleotide 1413) and mutant (A at nucleotide 1413) alleles were 0.56 and 0.44, respectively. The fourth pattern was found in only one FH heterozygote and was caused by heterozygosity at nucleotide 1469 (G/A). Nucleotide 1469 is the second base of codon 469Trp(TGG). The G→A mutation changes this codon into the amber stop codon, and is referred to as FH469→Stop. The mutant receptor consists of the amino terminal 468 amino acids. Because the truncated receptor has lost the membrane-spanning domain, it will not be anchored in the cell membrane. FH469→Stop destroys an AvaII restriction site, and this characteristic was used to develop a PCR method to establish its frequency in Norwegian FH subjects. Two out of 204 (1%) unrelated FH heterozygotes possessed the mutation.

Keywords

Polymerase Chain Reaction Codon Nonsense Mutation Familial Hypercholesterolemia Polymerase Chain Reaction Method 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References

  1. Davis CG, Goldstein JL, Südhof TC, Anderson RGW, Russell DW, Brown MS (1987) Acid-dependent ligand dissociation and recycling of LDL receptor mediated by growth factor homology region. Nature 326:760–765Google Scholar
  2. Funke H, Klug J, Frossard P, Coleman R, Assmann G (1986) PstI RFLP close to the LDL receptor gene. Nucleic Acids Res 14:7820Google Scholar
  3. Goldstein JL, Brown MS (1989) Familial hypercholesterolemia. In: Scriver CR, Baudet AL, Sly WS, Valle D (eds) The metabolic basis of inherited disease, 6th edn. McGraw-Hill, New York, pp 1215–1250Google Scholar
  4. Hayashi K (1991) PCR-SSCP: a simple and sensitive method for detection of mutations in the genomic DNA. PCR Methods Applic 1:34–38Google Scholar
  5. Hobbs HH, Esser V, Russell DW (1987) AvaII polymorphism in the human LDL receptor gene. Nucleic Acids Res 15:379Google Scholar
  6. Kotze MJ, Retief AE, Brink PA, Welch HFH (1986) A DNA polymorphism in the human low-density lipoprotein receptor gene. S Afr Med J 70:77–79Google Scholar
  7. Kotze MJ, Langenhoven E, Dietzsch E, Retief AE (1987) A RFLP associated with the low density lipoprotein receptor gene (LDLR). Nucleic Acids Res 15:376Google Scholar
  8. Leitersdorf E, Hobbs HH (1987) Human LDL receptor gene: two ApaLI RFLPs. Nucleic Acids Res 15:2782Google Scholar
  9. Leitersdorf E, Chakravarti A, Hobbs HH (1989) Polymorphic DNA haplotypes at the LDL receptor locus. Am J Hum Genet 44:409–421Google Scholar
  10. Leitersdorf E, Tobin EJ, Davignon J, Hobbs HH (1990) Common low-density lipoprotein receptor mutations in the French Canadian population. J Clin Invest 85:1014–1023Google Scholar
  11. Miyake Y, Tajima S, Funahashi T, Yamamoto A (1989) Analysis of a recycling-impaired mutant of low density lipoprotein receptor in familial hypercholesterolemia. J Biol Chem 264:16584–16590Google Scholar
  12. Müller C (1939) Angina pectoris in hereditary xanthomatosis. Arch Intern Med 64:675–700Google Scholar
  13. Orita M, Iwahana H, Kanazawa H, Hayashi K, Sekiya T (1989a) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc Natl Acad Sci USA: 86:2766–2770PubMedGoogle Scholar
  14. Orita M, Suzuki Y, Sekiya T, Hayashi K (1989b) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5:874–879PubMedGoogle Scholar
  15. Südhof TC, Goldstein JL, Brown MS, Russell DW (1985a) The LDL receptor gene: a mosaic of exons shared with different proteins. Science 228:815–822Google Scholar
  16. Südhof TC, Russell DW, Goldstein JL, Brown MS, Sanchcz-Pescado R, Bell GI (1985b) Cassette of eight exons shared by genes for LDL receptor and EGF precursor. Science 228:893–895Google Scholar
  17. Warnich L, Kotze MJ, Langenhoven E, Retief AE (1992) Detection of a frequent polymorphism in exon 10 of the low-density ipoprotein receptor gene. Hum Genet 89:362Google Scholar
  18. 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 39:27–38Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Trond P. Leren
    • 1
  • Kari Solberg
    • 1
    • 2
  • Olaug K. Rødningen
    • 1
    • 2
  • Oddveig Røsby
    • 1
  • Serena Tonstad
    • 3
  • Leiv Ose
    • 3
  • Kåre Berg
    • 1
    • 2
  1. 1.Department of Medical GeneticsUllevål University HospitalOsloNorway
  2. 2.Institute of Medical Genetics, University of OsloOsloNorway
  3. 3.Lipid Clinic, RikshospitaletOsloNorway

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