Ha V T, Marshall M K, Elsas L J, Pinnell S R, Yeowell H N
Division of Dermatology, Duke University Medical Center, Durham, North Carolina 27710.
J Clin Invest. 1994 Apr;93(4):1716-21. doi: 10.1172/JCI117155.
In the present study, we have isolated and sequenced the complementary DNAs of two mutant alleles for lysyl hydroxylase (LH) in fibroblasts from one patient (AT750) with Ehlers-Danlos syndrome type VI (EDS VI). We have identified a putative mutation in each allele which may be responsible for the patient's decreased LH (normalized to prolyl hydroxylase) activity (24% of normal). Intermediate levels of LH activity were measured in the patient's parents, who are clinically normal (father 52%; mother 86%). After the cloning of cDNAs and amplification by PCR, sequence analysis revealed two equally distributed populations of cDNAs for LH in the AT750 cell line. Each allele revealed different but significant changes from the normal sequence. In one allele (allele 1), the most striking change was a triple base deletion that would result in the loss of residue Glu532. The most significant difference in the other allele (allele 2) was a G-->A change which would produce a Gly678-->Arg codon change in a highly conserved region of the enzyme. Restriction analysis identified that allele 1 was inherited from the proband's mother and allele 2 from the father. This study represents the first example of compound heterozygosity for the LH gene in an EDS VI patient, and it appears that there is an additive effect of each mutant allele on clinical expression in this patient.
在本研究中,我们从一名患有Ⅵ型埃勒斯-当洛综合征(EDS VI)的患者(AT750)的成纤维细胞中分离并测序了赖氨酰羟化酶(LH)的两个突变等位基因的互补DNA。我们在每个等位基因中都鉴定出一个可能导致患者LH(相对于脯氨酰羟化酶标准化后)活性降低(为正常活性的24%)的推定突变。在临床正常的患者父母中检测到中等水平的LH活性(父亲为52%;母亲为86%)。在克隆cDNA并通过PCR扩增后,序列分析显示在AT750细胞系中存在两个等量分布的LH cDNA群体。每个等位基因与正常序列相比都有不同但显著的变化。在一个等位基因(等位基因1)中,最显著的变化是三个碱基的缺失,这将导致Glu532残基的丢失。另一个等位基因(等位基因2)中最显著的差异是一个G→A的变化,这将在该酶的一个高度保守区域产生Gly678→Arg密码子的改变。限制性分析确定等位基因1遗传自先证者的母亲,等位基因2遗传自父亲。本研究代表了EDS VI患者中LH基因复合杂合性的首个实例,并且似乎每个突变等位基因对该患者的临床表型都有累加效应。