Gache Y, Allegra M, Bodemer C, Pisani-Spadafora A, de Prost Y, Ortonne J P, Meneguzzi G
INSERM U385, Faculté de Médecine, 06107 Nice Cedex 2, France.
Hum Mol Genet. 2001 Oct 1;10(21):2453-61. doi: 10.1093/hmg/10.21.2453.
Change of the clinical picture with aging is noted in some patients suffering from junctional epidermolysis bullosa (JEB), an inherited blistering disorder caused by extensive disadhesion of the epithelia. We have studied a patient born with severe JEB associated with absent expression of laminin 5. A remarkable reduction of the blistering tendency was observed with aging that correlated with a restored expression of immunoreactive laminin 5 molecules. Genetic analysis of the gene LAMB3 detected compound heterozygosity for the nonsense mutation R635X and a novel 2 bp deletion (1587delAG) resulting in a downstream premature termination codon. RT-PCR amplification of total RNA purified from skin biopsies demonstrated that the mutated beta3 mRNAs underwent rapid decay shortly after birth, and that illegitimate splicing of the mRNA carrying mutation 1587delAG generated a new internally shortened beta3 transcript with advancing age. Our genetic and biochemical data show that (i) the illegitimate splicing of the beta3 pre-mRNA results in synthesis and secretion of a laminin 5 heterotrimer with an internally deleted beta3 polypeptide, (ii) expression of the mutated beta3 polypeptide is up-regulated in the basal keratinocytes with high proliferative potential, (iii) absence of the N-terminal region of the beta3 rod domain II thought to stabilize the tertiary structure of the laminin 5 is not required for the assembly of the protein and (iv) the mutant laminin 5 retains its adhesive potential. Our results demonstrate that mRNA rescue may underlie the evolution of the clinical phenotype in inherited skin conditions.
在一些患有交界性大疱性表皮松解症(JEB)的患者中,注意到临床症状会随着年龄增长而发生变化,JEB是一种由上皮细胞广泛分离引起的遗传性水疱性疾病。我们研究了一名出生时患有严重JEB且层粘连蛋白5表达缺失的患者。随着年龄增长,水疱形成倾向显著降低,这与免疫反应性层粘连蛋白5分子的表达恢复相关。对LAMB3基因的遗传分析检测到无义突变R635X和一个新的2bp缺失(1587delAG)的复合杂合性,导致下游出现提前终止密码子。从皮肤活检样本中纯化的总RNA进行RT-PCR扩增表明,突变的β3 mRNA在出生后不久迅速降解,并且携带突变1587delAG的mRNA的异常剪接随着年龄增长产生了一种新的内部缩短的β3转录本。我们的遗传和生化数据表明:(i)β3前体mRNA的异常剪接导致合成并分泌一种具有内部缺失β3多肽的层粘连蛋白5异源三聚体;(ii)突变的β3多肽在具有高增殖潜能的基底角质形成细胞中表达上调;(iii)认为对稳定层粘连蛋白5三级结构至关重要的β3杆状结构域II的N端区域缺失对于该蛋白的组装并非必需;(iv)突变的层粘连蛋白5保留了其黏附潜能。我们的结果表明,mRNA拯救可能是遗传性皮肤病临床表型演变的基础。