Takeda Haruko, Takami Marika, Oguni Tomoko, Tsuji Takehito, Yoneda Kazuhiro, Sato Hiroaki, Ihara Naoya, Itoh Tomohito, Kata Srinivas R, Mishina Yuji, Womack James E, Moritomo Yasuo, Sugimoto Yoshikazu, Kunieda Tetsuo
Shirakawa Institute of Animal Genetics, Nishi-shirakawa, Fukushima 961-8061, Japan.
Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10549-54. doi: 10.1073/pnas.152337899. Epub 2002 Jul 22.
Chondrodysplastic dwarfism in Japanese brown cattle is an autosomal recessive disorder characterized by short limbs. Previously, we mapped the locus responsible for the disease on the distal end of bovine chromosome 6. Here, we narrowed the critical region to approximately 2 cM by using linkage analysis, constructed a BAC and YAC contig covering this region, and identified a gene, LIMBIN (LBN), that possessed disease-specific mutations in the affected calves. One mutation was a single nucleotide substitution leading to an activation of a cryptic splicing donor site and the other was a one-base deletion resulting in a frameshift mutation. Strong expression of the Lbn gene was observed in limb buds of developing mouse embryos and in proliferating chondrocytes and bone-forming osteoblasts in long bones. These findings indicate that LBN is responsible for bovine chondrodysplastic dwarfism and has a critical role in a skeletal development.
日本褐牛的软骨发育不全侏儒症是一种以四肢短小为特征的常染色体隐性疾病。此前,我们将该疾病相关基因座定位到牛6号染色体的远端。在此,我们通过连锁分析将关键区域缩小至约2厘摩,构建了覆盖该区域的细菌人工染色体(BAC)和酵母人工染色体(YAC)重叠群,并鉴定出一个基因,即肢体结合蛋白(LIMBIN,LBN),该基因在患病犊牛中存在疾病特异性突变。一个突变是单核苷酸替换,导致一个隐蔽剪接供体位点激活,另一个突变是单碱基缺失,导致移码突变。在发育中小鼠胚胎的肢芽以及长骨中增殖的软骨细胞和成骨的成骨细胞中观察到Lbn基因的强表达。这些发现表明,LBN是牛软骨发育不全侏儒症的致病基因,并且在骨骼发育中起关键作用。