Kraft H J, Hendriks W, de Jong W W, Lubsen N H, Schoenmakers J G
Department of Molecular Biology, University of Nijmegen, The Netherlands.
J Mol Biol. 1993 Feb 20;229(4):849-59. doi: 10.1006/jmbi.1993.1091.
In duck the single copy lactate dehydrogenase B (LDH-B) gene also encodes an abundant lens protein, epsilon-crystallin. The LDH-B/epsilon-crystallin gene consists of eight exons, of which the first is non-coding. The promoter region lacks a TATA box, is very GC-rich and contains multiple consensus Sp1 binding sites. The gene has two discrete transcription start sites located 28 base-pairs apart. Both sites are used about equally in heart tissue, while transcription from the downstream start site predominates in the lens. For maximal promoter activity in lens or heart, sequences from the first intron are required. The enhancer(s) in this intron is promoter specific as it could not activate the tk promoter. Studies at the RNA level show that the overexpression of the LDH-B/epsilon-crystallin gene in the lens is regulated at the transcriptional level, yet no tissue-specific regulatory elements could be detected in a region spanning from -1.9 kb (1 kb = 10(3) bases or base-pairs) up to the translation initiation site in the second exon. The basis for the differential expression of the LDH-B/epsilon-crystallin gene in duck heart and lens is the usage of the downstream transcription initiation site. However, our results do not allow a distinction between activation of the downstream transcription start site in the lens or repression of the use of this site in heart.
在鸭中,单拷贝的乳酸脱氢酶B(LDH - B)基因还编码一种丰富的晶状体蛋白,即ε - 晶状体蛋白。LDH - B/ε - 晶状体蛋白基因由八个外显子组成,其中第一个外显子是非编码的。启动子区域缺乏TATA框,富含GC,并且包含多个共有Sp1结合位点。该基因有两个相距28个碱基对的离散转录起始位点。在心脏组织中,两个位点的使用频率大致相同,而在晶状体中,下游起始位点的转录占主导。为了在晶状体或心脏中获得最大启动子活性,需要第一个内含子的序列。该内含子中的增强子具有启动子特异性,因为它不能激活tk启动子。在RNA水平的研究表明,晶状体中LDH - B/ε - 晶状体蛋白基因的过表达在转录水平受到调控,但在从 - 1.9 kb(1 kb = 10³个碱基或碱基对)到第二个外显子中翻译起始位点的区域内,未检测到组织特异性调控元件。鸭心脏和晶状体中LDH - B/ε - 晶状体蛋白基因差异表达的基础是下游转录起始位点的使用情况。然而,我们的结果无法区分晶状体中下游转录起始位点的激活或心脏中该位点使用的抑制情况。