Spirito F, Chavanas S, Prost-Squarcioni C, Pulkkinen L, Fraitag S, Bodemer C, Ortonne J P, Meneguzzi G
U385 INSERM, Faculté de Médecine, Avenue de Valombrose, 06107 Nice cedex 2, France.
J Biol Chem. 2001 Jun 1;276(22):18828-35. doi: 10.1074/jbc.M100381200. Epub 2001 Mar 14.
Laminin 5, the major keratinocyte adhesion ligand, is found in the lamina lucida subregion of the epidermal basement membrane of the skin, where it colocalizes with the anchoring filaments. Mutations in the genes encoding laminin 5 cause junctional epidermolysis bullosa, an inherited skin blistering disease characterized by abnormal hemidesmosomes and cleavage of the lamina lucida leading to epidermal detachment. In this work we describe the genetic basis of a new subtype of lethal inherited epidermolysis bullosa associated with reduced skin reactivity to laminin 5, presence of mature hemidesmosomes, and intradermal cleavage of the skin. The epidermolysis bullosa patients were heterozygous for a nonsense mutation (Q896X) and a splice site mutation (764-10T-->G) in the gene (LAMC2) for the gamma2 chain of laminin 5. The nonsense mutation causes accelerated decay of the corresponding mRNA, while the splice site mutation results in maturation of a cryptic wild-type gamma2 mRNA leading to reduced expression of wild-type laminin 5. In vitro studies using the probands' keratinocytes showed that secretion of reduced amounts of functional laminin 5 in the patient, although permitting formation of hemidesmosomes, fail to restore efficient cell adhesion. Our results provide the first evidence that laminin 5 contributes to the firm adhesion of the epithelial basement membrane to the underlying stroma. They also show that a low expression level of laminin 5 induces assembly of mature hemidesmosomes in vivo but fails to assure a stable cohesion of the dermal-epidermal junction.
层粘连蛋白5是角质形成细胞的主要黏附配体,存在于皮肤表皮基底膜的透明层亚区域,在那里它与锚定细丝共定位。编码层粘连蛋白5的基因突变会导致交界性大疱性表皮松解症,这是一种遗传性皮肤水疱病,其特征是半桥粒异常以及透明层的裂解导致表皮脱离。在这项研究中,我们描述了一种致死性遗传性大疱性表皮松解症新亚型的遗传基础,该亚型与皮肤对层粘连蛋白5的反应性降低、成熟半桥粒的存在以及皮肤真皮内裂解有关。大疱性表皮松解症患者在层粘连蛋白5γ2链基因(LAMC2)中存在一个无义突变(Q896X)和一个剪接位点突变(764 - 10T→G),为杂合子。无义突变导致相应mRNA加速降解,而剪接位点突变导致一个隐蔽的野生型γ2 mRNA成熟,从而导致野生型层粘连蛋白5表达减少。使用先证者角质形成细胞进行的体外研究表明,患者分泌的功能性层粘连蛋白5量减少,尽管允许形成半桥粒,但未能恢复有效的细胞黏附。我们的结果首次证明层粘连蛋白5有助于上皮基底膜与下方基质的牢固黏附。它们还表明,层粘连蛋白5的低表达水平在体内诱导成熟半桥粒的组装,但不能确保真皮 - 表皮连接的稳定黏附。