Imaging cellular responses to antigen tagged DNA damage.

Imaging cellular responses to antigen tagged DNA damage.
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成像细胞对抗原标记 DNA 损伤的反应

DOI:
10.1016/j.dnarep.2018.08.023
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发表时间:
2018-11
期刊:
影响因子:
3.8
通讯作者:
Seidman MM
Seidman MM
中科院分区:
医学3区
文献类型:
--
作者:
Bellani MA;Huang J;Paramasivam M;Pokharel D;Gichimu J;Zhang J;Seidman MM

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由于许多研究者数十年来出色的生物化学研究,共价DNA损伤的修复途径得到了相当详细的了解。这些实验的一个重要特征是与纯化蛋白或无细胞提取物一起孵育的寡核苷酸或质粒底物上的确定加合物位置。除了一些例外,当研究转向活的哺乳动物细胞对基因组DNA中相同加合物的反应时,这种确定性就失去了。这反映了测定的局限性,例如基于免疫荧光的测定,其广泛用于追踪暴露于DNA反应性化合物的细胞中的响应蛋白。缺乏有效的加合物检测试剂意味着必须假设反应蛋白和加合物之间的接近性。由于这些假设可能不正确,基于体外系统的模型可能无法解释体内观察结果。在这里,我们讨论了使用检测标签来解决病变位置的问题,如我们最近的工作所示的复制依赖和独立的反应链间交联。
Repair pathways of covalent DNA damage are understood in considerable detail due to decades of brilliant biochemical studies by many investigators. An important feature of these experiments is the defined adduct location on oligonucleotide or plasmid substrates that are incubated with purified proteins or cell free extracts. With some exceptions, this certainty is lost when the inquiry shifts to the response of living mammalian cells to the same adducts in genomic DNA. This reflects the limitation of assays, such as those based on immunofluorescence, that are widely used to follow responding proteins in cells exposed to a DNA reactive compound. The lack of effective reagents for adduct detection means that the proximity between responding proteins and an adduct must be assumed. Since these assumptions can be incorrect, models based on in vitro systems may fail to account for observations made in vivo. Here we discuss the use of a detection tag to address the problem of lesion location, as illustrated by our recent work on replication dependent and independent responses to interstrand crosslinks.
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