利用果蝇遗传学鉴定体内影响 PARP1 活性的因素。

Using Drosophila Genetics to Identify Factors that Affect PARP1 Activity In Vivo.

机构信息

Department of Biomedical Sciences, School of Medicine and Health Sciences, University of North Dakota, Grand Forks, ND, USA.

出版信息

Methods Mol Biol. 2023;2609:339-352. doi: 10.1007/978-1-0716-2891-1_20.

DOI:10.1007/978-1-0716-2891-1_20

PMID:36515845

Abstract

PARP1 is the enzyme responsible for the majority of the poly(ADP-ribose) (pADPr) synthesis in Drosophila. Its activity can be easily evaluated in vitro by measuring the level of pADPr, which allow to study the effect of potential PARP1 upstream factors on PARP1 activity. However, PARP1 activity can be challenging to measure in vivo, due to the presence of PARG, since pADPr level is a consequence of the activity of both PARP1 that synthetizes pADPr and PARG that degrades it. An increase in PARG activity can hide an increase of PARP1 activity. In this context, the effect of potential upstream factors on PARP1 activity can be hard to measure. Here, we describe a genetic background where PARG is absent to study changes in PARP1 activity at different developmental time points.

摘要

PARP1 是负责在果蝇中合成大部分聚（ADP-核糖）（pADPr）的酶。其活性可以通过测量 pADPr 的水平在体外轻松评估，这使得可以研究潜在的 PARP1 上游因素对 PARP1 活性的影响。然而，由于 PARG 的存在，PARP1 活性在体内很难测量，因为 pADPr 水平是 PARP1 合成 pADPr 和 PARG 降解它的活性的结果。PARG 活性的增加可能会掩盖 PARP1 活性的增加。在这种情况下，潜在的上游因素对 PARP1 活性的影响很难测量。在这里，我们描述了一个缺乏 PARG 的遗传背景，以研究不同发育时间点 PARP1 活性的变化。