Positioning effects of KillerRed inside of cells correlate with DNA strand breaks after activation with visible light.
Positioning effects of KillerRed inside of cells correlate with DNA strand breaks after activation with visible light.
复制标题
细胞内部的Killerr的定位效应与可见光激活后与DNA链断裂相关。
DOI：
10.7150/ijms.8.97
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发表时间：
2011-01-21
影响因子：
3.6
通讯作者：
Braun K
中科院分区：
文献类型：
--
作者：
Waldeck W;Mueller G;Wiessler M;Tóth K;Braun K
荧光蛋白 (FP) 是用于新应用的既定工具，不仅限于细胞生物学研究。它们在手术中也可能是理想的选择，可以提高区分目标组织和周围健康组织的精度。这里使用的像 KillerRed (KRED) 这样的 FP 可以通过可见日光激发来激活，发射活性电子，产生活性氧 (ROS)，从而产生光杀伤过程。可以肯定的是，KRED 的细胞毒性程度取决于其亚细胞定位。有证据表明，核纤层，尤其是染色质，是 KRED 介导的基于 ROS 的 DNA 损伤的关键目标。在这里，我们研究了与核纤层和组蛋白 H2A DNA 结合蛋白融合的 KRED 蛋白的破坏作用。我们检测到 DNA 链断裂的频率，首先取决于照明时间，其次取决于染色质定位与 ROS 产生位点之间的空间距离。因此，我们可以识别出 200、400 和 (600) bp 的明确 DNA 带作为最显着的降解产物，大概代表 KRED 诱导的核小体间 DNA 裂解。这些发现不仅限于PDT等治疗领域中程序性细胞死亡过程的检测，而且还有助于更好地理解表观基因组世界中的结构-功能关系。
Fluorescent proteins (FPs) are established tools for new applications, not-restricted to the cell biological research. They could also be ideal in surgery enhancing the precision to differentiate between the target tissue and the surrounding healthy tissue. FPs like the KillerRed (KRED), used here, can be activated by excitation with visible day-light for emitting active electrons which produce reactive oxygen species (ROS) resulting in photokilling processes. It is a given that the extent of the KRED's cell toxicity depends on its subcellular localization. Evidences are documented that the nuclear lamina as well as especially the chromatin are critical targets for KRED-mediated ROS-based DNA damaging. Here we investigated the damaging effects of the KRED protein fused to the nuclear lamina and to the histone H2A DNA-binding protein. We detected a frequency of DNA strand breaks, dependent first on the illumination time, and second on the spatial distance between the localization at the chromatin and the site of ROS production. As a consequence we could identify defined DNA bands with 200, 400 and (600) bps as most prominent degradation products, presumably representing an internucleosomal DNA cleavage induced by KRED. These findings are not restricted to the detection of programmed cell death processes in the therapeutic field like PDT, but they can also contribute to a better understanding of the structure-function relations in the epigenomic world.
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影响因子：
7.4
作者：
Circu, Magdalena L.;Aw, Tak Yee
通讯作者：
Aw, Tak Yee
DOI：
10.1615/critreveukargeneexpr.v9.i3-4.130
发表时间：
1999-01-01
影响因子：
1.6
作者：
Goldberg, M;Harel, A;Gruenbaum, Y
通讯作者：
Gruenbaum, Y
影响因子：
7.5
作者：
Cremer, Thomas;Cremer, Marion;Fakan, Stanislav
通讯作者：
Fakan, Stanislav
影响因子：
4.3
作者：
Chatterjee, S;Berger, SJ;Berger, NA
通讯作者：
Berger, NA
影响因子：
11.5
作者：
Donawho, Cherrie K.;Luo, Yan;Frost, David J.
通讯作者：
Frost, David J.