Cell array-based intracellular localization screening reveals novel functional features of human chromosome 21 proteins.
Cell array-based intracellular localization screening reveals novel functional features of human chromosome 21 proteins.
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DOI：
10.1186/1471-2164-7-155
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发表时间：
2006-06-16
期刊：
影响因子：
4.4
通讯作者：
Janitz M
中科院分区：
文献类型：
--
作者：
Hu YH;Warnatz HJ;Vanhecke D;Wagner F;Fiebitz A;Thamm S;Kahlem P;Lehrach H;Yaspo ML;Janitz M
人类21号染色体三体导致唐氏综合症，这是一种复杂的发育和神经退行性疾病。然而，对唐氏综合症的分子分析提出了一个特别的挑战，因为chr21的非整倍体区域包含许多未知功能的基因。对人类chr21蛋白的亚细胞定位有助于进一步了解编码这些蛋白的基因的功能和调控机制。根据这一想法，我们使用了一种转基因细胞阵列技术，对Chr 21蛋白在细胞内的分布进行了快速和经济有效的分析。我们选择了分布在大多数21q上的89个基因，范围从RBM11(14.5Mb)到MCM3AP(46.6Mb)，其中部分基因在唐氏综合征小鼠模型中异常表达。将这些基因的开放阅读框克隆到带有氨基末端His6标签的哺乳动物表达载体中。将构建好的重组载体排列在玻片上，反转染HEK293T细胞进行蛋白表达。使用一组细胞器标记对每个chr21蛋白进行共定位检测。在这里，我们报告了52个蛋白质的亚细胞定位特性。对于其中的34种蛋白质，它们的定位是第一次被描述。此外，由于claudin-8和claudin-14基因的蛋白过表达而导致的细胞形态和生长的改变已经被表征。基于细胞阵列的蛋白质表达和检测方法是进行大规模功能分析的经济有效的平台，包括蛋白质亚细胞定位和细胞表型筛选。这一研究结果揭示了人类chr21蛋白的新功能特征，这将有助于进一步了解唐氏综合征的分子病理学。
Trisomy of human chromosome 21 (Chr21) results in Down's syndrome, a complex developmental and neurodegenerative disease. Molecular analysis of Down's syndrome, however, poses a particular challenge, because the aneuploid region of Chr21 contains many genes of unknown function. Subcellular localization of human Chr21 proteins may contribute to further understanding of the functions and regulatory mechanisms of the genes that code for these proteins. Following this idea, we used a transfected-cell array technique to perform a rapid and cost-effective analysis of the intracellular distribution of Chr 21 proteins. We chose 89 genes that were distributed over the majority of 21q, ranging from RBM11 (14.5 Mb) to MCM3AP (46.6 Mb), with part of them expressed aberrantly in the Down's syndrome mouse model. Open reading frames of these genes were cloned into a mammalian expression vector with an amino-terminal His6 tag. All of the constructs were arrayed on glass slides and reverse transfected into HEK293T cells for protein expression. Co-localization detection using a set of organelle markers was carried out for each Chr21 protein. Here, we report the subcellular localization properties of 52 proteins. For 34 of these proteins, their localization is described for the first time. Furthermore, the alteration in cell morphology and growth as a result of protein over-expression for claudin-8 and claudin-14 genes has been characterized. The cell array-based protein expression and detection approach is a cost-effective platform for large-scale functional analyses, including protein subcellular localization and cell phenotype screening. The results from this study reveal novel functional features of human Chr21 proteins, which should contribute to further understanding of the molecular pathology of Down's syndrome.
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影响因子：
64.5
作者：
Cawley, S;Bekiranov, S;Gingeras, TR
通讯作者：
Gingeras, TR
影响因子：
7.7
作者：
Simpson, JC;Wellenreuther, R;Wiemann, S
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Wiemann, S
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作者：
Accola, MA;Huang, B;McNiven, MA
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McNiven, MA
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作者：
Liebel, U;Starkuviene, V;Pepperkok, R
通讯作者：
Pepperkok, R
DOI：
10.1152/ajplung.00423.2003
发表时间：
2004-12-01
影响因子：
4.9
作者：
Daugherty, BL;Mateescu, M;Koval, M
通讯作者：
Koval, M