A Mutation in the Human Glucocerebrosidase Gene in Neuronopathic Gaucher's Disease

Abstract

To search for a genetic marker for type 2 Gaucher's disease (acute neuronopathic form), we compared the nucleotide sequence of a cloned glucocerebrosidase gene from a patient with Gaucher's disease with a normal gene. We found only a single base substitution (T----C) in exon X. This mutation results in the substitution of proline for leucine in position number 444 and produces a new cleavage site for the NciI restriction endonuclease. We analyzed NciI enzymatic digests of genomic DNA from 20 patients with type 1, 5 with type 2, and 11 with type 3 Gaucher's disease, and 29 normal controls for a restriction-fragment-length polymorphism (RFLP). Four of 5 patients with type 2 disease and all 11 with type 3 disease had at least one allele with the mutation. Two of 5 patients with type 2 disease and 7 of 11 with type 3 were homozygous for this mutation. Only 4 of 20 patients with type 1 Gaucher's disease had the mutant allele and were heterozygous for it. None of the 29 normal controls had the mutant allele. The high frequency of this mutation (444leucine----proline) in patients with neuronopathic Gaucher's disease, detectable by the NciI RFLP, may be of value in the identification of patients who will have the neurologic sequelae of Gaucher's disease.

Gaucher disease: mutation and polymorphism spectrum in the glucocerebrosidase gene (GBA)

“…Following the current recommendation that all variants be described at the most basic level (Human Genome Variation Society; www.hgvs.org/mutnomen), mutations discussed in the text are referred to by cDNA nomenclature, again with the traditional name of the allele included in parentheses to aid in recognition. The first two GBA mutations were described in the late 1980s [Tsuji et al, 1987[Tsuji et al, , 1988. These two alleles, c.1448T4C (L444P) and c.1226A4G (N370S), are still the prevalent mutant alleles encountered in most populations.…”

“…It was widely anticipated that the elucidation of the GBA sequence would yield an explanation of the vast phenotypic differences encountered among patients with GD. During the past two decades, there have been significant advances in understanding the organization of the gene and locus [Winfield et al, 1997], the identification of mutant alleles [Beutler and Gelbart, 1993;Grabowski and Horowitz, 1997;Tsuji et al, 1987], the enzyme structure [Dvir et al, 2003], and genotype-phenotype correlations [Beutler, 1992;Grabowski, 1997;Koprivica et al, 2000;Sidransky et al, 1994;Zimran et al, 1989]. Nevertheless, the pathological mechanisms through which the mutant enzyme acts to cause the diverse range of patient phenotypes are still elusive.…”

Genetic heterogeneity in type 1 Gaucher disease: Multiple genotypes in Ashkenazic and non-Ashkenazic individuals

“…In contrast, 15 type 1 patients had one allele with the mutation, and 3 additional patients were homozygous for the mutation. Interestingly, 1 of the type 1 patients had one allele with the Asn-370 to Ser mutation and another allele with the Leu-444 to Pro mutation that had been described (13) in patients with neuronopathic Gaucher disease.…”

“…High molecular weight DNA was prepared from autopsy or biopsy tissues, mononuclear blood cells, or cultured skin fibroblasts as described (13).…”

“…A genomic library was constructed from an Ashkenazic Jewish patient with type 1 Gaucher disease. The procedures for the construction of the genomic library with DNA isolated from cultured skin fibroblasts as well as the cloning of the glucocerebrosidase gene have been described in detail elsewhere (13,14). The BamHI fragments of the glucocerebrosidase genomic DNA were subcloned into pUC19.…”