Tayebi Nahid, Stubblefield Barbara K, Park Joseph K, Orvisky Eduard, Walker Jamie M, LaMarca Mary E, Sidransky Ellen
Clinical Neuroscience Branch, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892, USA.
Am J Hum Genet. 2003 Mar;72(3):519-34. doi: 10.1086/367850. Epub 2003 Feb 13.
Gaucher disease results from an autosomal recessive deficiency of the lysosomal enzyme glucocerebrosidase. The glucocerebrosidase gene is located in a gene-rich region of 1q21 that contains six genes and two pseudogenes within 75 kb. The presence of contiguous, highly homologous pseudogenes for both glucocerebrosidase and metaxin at the locus increases the likelihood of DNA rearrangements in this region. These recombinations can complicate genotyping in patients with Gaucher disease and contribute to the difficulty in interpreting genotype-phenotype correlations in this disorder. In the present study, DNA samples from 240 patients with Gaucher disease were examined using several complementary approaches to identify and characterize recombinant alleles, including direct sequencing, long-template polymerase chain reaction, polymorphic microsatellite repeats, and Southern blots. Among the 480 alleles studied, 59 recombinant alleles were identified, including 34 gene conversions, 18 fusions, and 7 downstream duplications. Twenty-two percent of the patients evaluated had at least one recombinant allele. Twenty-six recombinant alleles were found among 310 alleles from patients with type 1 disease, 18 among 74 alleles from patients with type 2 disease, and 15 among 96 alleles from patients with type 3 disease. Several patients carried two recombinations or mutations on the same allele. Generally, alleles resulting from nonreciprocal recombination (gene conversion) could be distinguished from those arising by reciprocal recombination (crossover and exchange), and the length of the converted sequence was determined. Homozygosity for a recombinant allele was associated with early lethality. Ten different sites of crossover and a shared pentamer motif sequence (CACCA) that could be a hotspot for recombination were identified. These findings contribute to a better understanding of genotype-phenotype relationships in Gaucher disease and may provide insights into the mechanisms of DNA rearrangement in other disorders.
戈谢病是由于溶酶体酶葡萄糖脑苷脂酶的常染色体隐性缺乏所致。葡萄糖脑苷脂酶基因位于1q21的一个基因丰富区域,该区域在75kb范围内包含六个基因和两个假基因。该位点同时存在葡萄糖脑苷脂酶和亚细胞定位蛋白的相邻、高度同源假基因,增加了该区域DNA重排的可能性。这些重组会使戈谢病患者的基因分型复杂化,并导致难以解释该疾病的基因型-表型相关性。在本研究中,使用几种互补方法对240例戈谢病患者的DNA样本进行了检测,以鉴定和表征重组等位基因,包括直接测序、长模板聚合酶链反应、多态性微卫星重复和Southern印迹。在所研究的480个等位基因中,鉴定出59个重组等位基因,包括34个基因转换、18个融合和7个下游重复。22%的评估患者至少有一个重组等位基因。在1型疾病患者的310个等位基因中发现26个重组等位基因,2型疾病患者的74个等位基因中有18个,3型疾病患者的96个等位基因中有15个。几名患者在同一等位基因上携带两种重组或突变。一般来说,非相互重组(基因转换)产生的等位基因可以与相互重组(交叉和交换)产生的等位基因区分开来,并确定了转换序列的长度。重组等位基因的纯合性与早期致死率相关。鉴定出10个不同的交叉位点和一个可能是重组热点区的共享五聚体基序序列(CACCA)。这些发现有助于更好地理解戈谢病的基因型-表型关系,并可能为其他疾病中DNA重排的机制提供见解。