Lok S, Stevens W, Breitman M L, Tsui L C
Department of Genetics, Hospital for Sick Children, Toronto, Ontario, Canada.
Nucleic Acids Res. 1989 May 11;17(9):3563-82. doi: 10.1093/nar/17.9.3563.
Crystallins are the major water-soluble proteins of the vertebrate eye lens. These lens-specific proteins are encoded by several multi-gene families whose expression is differentially regulated during development. Our previous studies showed that the mouse gamma 2-crystallin promoter is active on transfection into lens-explant cultures derived from 14-day-old chick embryos but not on transfection into a variety of non-lens cells. In this study, transient expression data show that a sequence of 226 nucleotides upstream from the transcription start site is sufficient for activity of this promoter in the chicken lens cells. This sequence can be further divided into two domains, A and B, both of which are required for promoter function. Domain A (nucleotide -68 to -18) contains the TATA box and sequence motifs that are conserved in all gamma-crystallin promoters. Domain B (-226 to -120) consists of three regions. One of these regions contains an element with dyad symmetry and a sequence similar to the octamer motif. The second region contains an enhancer core consensus sequence. Two "enhancer-like" activities have been detected, one in Domain B and a second in a more distal region (-392 to -278) that does not appear to be required for promoter activity in transfection assays.
晶状体蛋白是脊椎动物眼晶状体中的主要水溶性蛋白质。这些晶状体特异性蛋白质由几个多基因家族编码,其表达在发育过程中受到差异调节。我们之前的研究表明,小鼠γ2-晶状体蛋白启动子在转染到源自14日龄鸡胚的晶状体外植体培养物中时具有活性,但在转染到多种非晶状体细胞中时则没有活性。在本研究中,瞬时表达数据表明,转录起始位点上游226个核苷酸的序列足以使该启动子在鸡晶状体细胞中发挥活性。该序列可进一步分为A和B两个结构域,两者都是启动子功能所必需的。结构域A(核苷酸-68至-18)包含TATA盒和在所有γ-晶状体蛋白启动子中保守的序列基序。结构域B(-226至-120)由三个区域组成。其中一个区域包含一个具有二元对称性的元件和一个与八聚体基序相似的序列。第二个区域包含一个增强子核心共有序列。已检测到两种“增强子样”活性,一种在结构域B中,另一种在更远端的区域(-392至-278),在转染实验中该区域似乎不是启动子活性所必需的。