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夜蛋白的功能分析:通过差异显示鉴定的一种生物钟调节基因

Functional analysis of nocturnin: a circadian clock-regulated gene identified by differential display.

作者信息

Baggs Julie E, Green Carla B

机构信息

Department of Biology, Center for Biological Timing, University of Virginia, Charlottesville, USA.

出版信息

Methods Mol Biol. 2006;317:243-54. doi: 10.1385/1-59259-968-0:243.

Abstract

Within the retina there is a circadian clock that controls the 24-h timing of processes such as hormone release, cell movement, and gene transcription. In an effort to better understand the molecular nature of this retinal clock, a differential display (DD) screen was performed to isolate a gene with high amplitude circadian rhythmicity in the Xenopus retina. A novel gene expressed in the early evening in photoreceptor cells was isolated and named nocturnin for night factor. This article outlines the steps we took to study a protein of unknown function, particularly highlighting the analyses one can perform when little more than the primary sequence of a gene is known. In addition, we describe the results of sequence analysis that assisted in predicting the function of nocturnin. We have shown that nocturnin acts as a deadenylase in vitro, removing the poly(A) tail from a mature messenger RNA in a process that either leads to degradation or translational silencing of a message. Although the role of nocturnin in the retina is unknown, future studies to identify target mRNAs that are deadenylated by nocturnin will assist in elucidating its physiological role in this tissue.

摘要

视网膜内存在一个昼夜节律时钟,它控制着诸如激素释放、细胞运动和基因转录等过程的24小时定时。为了更好地理解这个视网膜时钟的分子本质,我们进行了差异显示(DD)筛选,以在非洲爪蟾视网膜中分离出一个具有高振幅昼夜节律性的基因。我们分离出了一个在傍晚早期在光感受器细胞中表达的新基因,并将其命名为nocturnin(夜间因子)。本文概述了我们研究一种功能未知蛋白质所采取的步骤,特别强调了在仅知道基因的一级序列时可以进行的分析。此外,我们描述了有助于预测nocturnin功能的序列分析结果。我们已经表明,nocturnin在体外作为一种去腺苷酸化酶,在一个导致信使RNA降解或翻译沉默的过程中从成熟信使RNA上去除多聚(A)尾巴。尽管nocturnin在视网膜中的作用尚不清楚,但未来鉴定被nocturnin去腺苷酸化的靶标mRNA的研究将有助于阐明其在该组织中的生理作用。

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