Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events.

Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events.
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盘基网柄菌作为评估位点特异性 ADP-核糖基化事件的模型。

DOI:
10.1007/978-1-4939-8588-3_9
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发表时间:
2018
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
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通讯作者:
Kolb AL
Kolb AL
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作者:
Kolb AL

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盘状网柄阿米巴是一种单细胞生物,可以经历一个简单的发育程序,使其成为研究细胞运动,信号转导和细胞类型分化的分子机制的极好模型。在这种生物体中发现了许多在其他无脊椎动物模型中缺失或显示出有限保守性的人类基因。这包括ADP-核糖基转移酶,也称为聚ADP-核糖聚合酶(PARP),其是催化将单个或聚ADP-核糖部分添加到靶蛋白上的蛋白质家族。网柄菌的遗传易处理性及其相对简单的基因组结构使得它有可能破坏PARP基因组合,以及内源基因座上的特定ADP-核糖基化位点。总之,这使得Dictyosteliuman有吸引力的模型来评估ADP-核糖基化如何调节各种细胞过程,包括DNA修复,转录和细胞类型的规范。在这里,我们描述了一系列的技术来研究ADP-核糖基化inDictyosteopathy,包括ADP-核糖基化事件在体外和体内的分析,除了方法来评估这种修饰在体内的功能作用。
The amoebaDictyostelium discoideumis a single-cell organism that can undergo a simple developmental program, making it an excellent model to study the molecular mechanisms of cell motility, signal transduction, and cell-type differentiation. A variety of human genes that are absent or show limited conservation in other invertebrate models have been identified in this organism. This includes ADP-ribosyltransferases, also known as poly-ADP-ribose polymerases (PARPs), a family of proteins that catalyze the addition of single or poly-ADP-ribose moieties onto target proteins. The genetic tractability ofDictyosteliumand its relatively simple genome structure makes it possible to disrupt PARP gene combinations, in addition to specific ADP-ribosylation sites at endogenous loci. Together, this makesDictyosteliuman attractive model to assess how ADP-ribosylation regulates a variety of cellular processes including DNA repair, transcription, and cell-type specification. Here we describe a range of techniques to study ADP-ribosylation inDictyostelium, including analysis of ADP-ribosylation events in vitro and in vivo, in addition to approaches to assess the functional roles of this modification in vivo.
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