首页> 外文学位 >Outer segment localization signal at the C terminus of the photoreceptor-specific retinol dehydrogenase and proximal and distal sequences control UV cone pigment gene expression in transgenic zebrafish.
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Outer segment localization signal at the C terminus of the photoreceptor-specific retinol dehydrogenase and proximal and distal sequences control UV cone pigment gene expression in transgenic zebrafish.

机译:感光器特异性视黄醇脱氢酶C末端的外段定位信号以及近端和远端序列控制转基因斑马鱼中UV锥色素基因的表达。

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摘要

Photoreceptors are highly polarized neurons, and they have a very specialized structure, the outer segment (OS), responsible for light detection. Rhodopsin and other phototransduction proteins specifically localize there. Since all of the protein synthesis machinery is in the inner segment, there must be a very active OS transportation pathway.; Photoreceptor retinol dehydrogenase (prRDH) is a membrane-associated cytosolic protein in the OS of rods and cones. In this work, we found that the C-terminal 16 amino acids of prRDH confer membrane association as well as cone and rod OS targeting. Membrane association in transfected 293 cells and in transgenic Xenopus photoreceptors is mediated by fatty acylation at one or more evolutionarily conserved cysteines within the prRDH C-terminal tail. In bovine OS, native prRDH is similarly acylated. Efficient OS localization requires both membrane association and the prRDH sequence...(V/I)XPX at the extreme C-terminus, which closely resembles the C-terminal sequence that targets opsin/rhodopsin to the OS. Taken together, these data imply that the C-terminal...(V/I)XPX sequence is a general OS localization signal that can function in the context of both integral and peripheral membrane proteins.; Cones are responsible for color vision. In general, only one cone opsin gene is chosen to be expressed in a given cone. However, the molecular basis of cone photoreceptor-specific gene expression is largely unknown. In this work, we defined cis-acting DNA sequences that control the cell-type specific expression of the zebrafish ultraviolet (UV) cone pigment gene by transient expression of green fluorescent protein (GFP) in transgenic zebrafish embryos. These experiments show that 4.8 kb of 5' flanking sequences from the zebrafish UV pigment gene direct expression specifically to UV cones and that this activity requires both distal and proximal sequences. In addition, we demonstrate that a UV promoter proximal region located between -215 to -110 bp (with respect to the initiator methionine codon) can function in the context of a zebrafish rhodopsin promotor to convert its specificity from rod-only expression to rod and UV cone expression.
机译:感光细胞是高度极化的神经元,它们具有非常特殊的结构,即外部部分(OS),负责光检测。视紫红质和其他光转导蛋白特异性地定位在那里。由于所有蛋白质合成机制都在内部,因此必须有非常活跃的OS转运途径。感光视黄醇脱氢酶(prRDH)是视杆和视锥细胞OS中的膜相关胞质蛋白。在这项工作中,我们发现prRDH的C端16个氨基酸赋予膜结合以及视锥和视杆OS定位。在prRDH C末端尾部的一个或多个进化保守的半胱氨酸上,脂肪酰化介导了转染的293细胞和转基因非洲爪蟾感光细胞中的膜缔合。在牛OS中,天然prRDH同样被酰化。有效的OS定位既需要膜结合又需要prRDH序列...(V / I)XPX在极端C端,这非常类似于将视蛋白/视紫红质靶向OS的C端序列。综上所述,这些数据表明C末端...(V / I)XPX序列是一种通用的OS定位信号,可以在整合膜蛋白和外周膜蛋白的背景下发挥作用。锥体负责色觉。通常,仅选择一个视锥蛋白基因在给定视锥中表达。但是,锥感光细胞特异性基因表达的分子基础是很大程度上未知。在这项工作中,我们定义了顺式作用DNA序列,该序列通过在转基因斑马鱼胚胎中瞬时表达绿色荧光蛋白(GFP)来控制斑马鱼紫外线(UV)锥色素基因的细胞类型特异性表达。这些实验表明,来自斑马鱼UV色素基因的4.8 kb 5'侧翼序列特异性地直接表达至UV视锥,并且这种活性需要远端和近端序列。此外,我们证明了位于-215至-110 bp之间的UV启动子近端区域(相对于引发剂甲硫氨酸密码子)可以在斑马鱼视紫红质启动子的背景下发挥功能,将其特异性从仅棒表达转化为棒和UV锥体表达。

著录项

  • 作者

    Luo, Wenqin.;

  • 作者单位

    The Johns Hopkins University.;

  • 授予单位 The Johns Hopkins University.;
  • 学科 Biology Neuroscience.; Biology Molecular.; Biology Cell.
  • 学位 Ph.D.
  • 年度 2005
  • 页码 96 p.
  • 总页数 96
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 神经科学;分子遗传学;细胞生物学;
  • 关键词

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