McDermott J B, Peterson C A, Piatigorsky J
Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, MD 20892.
Gene. 1992 Aug 15;117(2):193-200. doi: 10.1016/0378-1119(92)90729-9.
The beta A1- and beta A3-crystallins are major polypeptides in the lenses of vertebrates. We present evidence that a single beta A3/A1 gene encodes these two proteins in the chicken. The beta A3/A1 gene has been sequenced and its functional promoter identified in transfection experiments. The chicken beta A3/A1 gene has the same structure as the human orthologue: six exons with standard splice sites and two alternative start codons from which the protein products are apparently translated. Northern analysis revealed an abundant 0.9-kb transcript in the lenses of 1-2-day-old chickens and no detectable transcripts in the rest of the eye, brain, heart, kidney, liver or skeletal muscle. The 5'-flanking sequence of the chicken beta A3/A1 gene is very similar to that of the human and mouse genes, suggesting conservation of important putative regulatory sequences in addition to the TATA box. A thymidine-rich element (bp -218 to -163) and a potential AP-1-binding site (bp -264 to -258) are present within the chicken 5'-flanking region. A DNA fragment from -382 to +22 of the chicken beta A3/A1 gene is sufficient to promote expression of the bacterial cat gene in transfected chicken primary lens epithelial cells, but not in transfected dermal fibroblasts.(ABSTRACT TRUNCATED AT 250 WORDS)
βA1-晶体蛋白和βA3-晶体蛋白是脊椎动物晶状体中的主要多肽。我们提供证据表明,鸡体内单个βA3/A1基因编码这两种蛋白质。βA3/A1基因已被测序,其功能启动子在转染实验中得以确定。鸡的βA3/A1基因与人类同源基因结构相同:有六个具有标准剪接位点的外显子以及两个可供选择的起始密码子,蛋白质产物显然是从这两个起始密码子开始翻译的。Northern分析显示,1至2日龄鸡的晶状体中有丰富的0.9 kb转录本,而在眼睛、大脑、心脏、肾脏、肝脏或骨骼肌的其他部位未检测到转录本。鸡βA3/A1基因的5'侧翼序列与人类和小鼠基因的序列非常相似,这表明除了TATA盒外,重要的假定调控序列也具有保守性。鸡5'侧翼区域内存在一个富含胸腺嘧啶的元件(碱基对-218至-163)和一个潜在的AP-1结合位点(碱基对-264至-258)。鸡βA3/A1基因从-382到+22的DNA片段足以促进细菌cat基因在转染的鸡原代晶状体上皮细胞中表达,但在转染的皮肤成纤维细胞中则不能。(摘要截短于250字)