Institute of Industrial Nanomaterials, Kumamoto University, Kumamoto 860-8555, Japan.
Faculty of Advanced Science and Technology, Kumamoto University, Kumamoto 860-8555, Japan.
Int J Mol Sci. 2024 Oct 3;25(19):10681. doi: 10.3390/ijms251910681.
DNA topoisomerase II (TOP2) is an enzyme that resolves DNA topological problems arising in various nuclear processes, such as transcription. Aclarubicin, a member of the anthracyclines, is known to prevent the association of TOP2 with DNA, inhibiting the early step of TOP2 catalytic reactions. During our research on the subnuclear distribution of human TOP2B, we found that aclarubicin affects the mobility of TOP2B in the nucleus. FRAP analysis demonstrated that aclarubicin decreased the nuclear mobility of EGFP-tagged TOP2B in a concentration-dependent manner. Aclarubicin exerted its inhibitory effects independently of TOP2B enzymatic activities: TOP2B mutants defective for either ATPase or topoisomerase activity also exhibited reduced nuclear mobility in the presence of aclarubicin. Immunofluorescence analysis showed that aclarubicin antagonized the induction of DNA damage by etoposide. Although the prevention of the TOP2-DNA association is generally considered a primary action of aclarubicin in TOP2 inhibition, our findings highlight a previously unanticipated effect of aclarubicin on TOP2B in the cellular environment.
DNA 拓扑异构酶 II(TOP2)是一种酶,可解决各种核过程中出现的 DNA 拓扑问题,如转录。阿克拉霉素是蒽环类药物的一种,已知可阻止 TOP2 与 DNA 的结合,从而抑制 TOP2 催化反应的早期步骤。在研究人类 TOP2B 的亚核分布时,我们发现阿克拉霉素会影响 TOP2B 在核内的迁移能力。FRAP 分析表明,阿克拉霉素以浓度依赖的方式降低了 EGFP 标记的 TOP2B 的核迁移能力。阿克拉霉素的抑制作用独立于 TOP2B 的酶活性:ATP 酶或拓扑异构酶活性有缺陷的 TOP2B 突变体在阿克拉霉素存在的情况下也表现出核迁移能力降低。免疫荧光分析表明,阿克拉霉素拮抗依托泊苷诱导的 DNA 损伤。虽然阻止 TOP2-DNA 结合通常被认为是阿克拉霉素抑制 TOP2 的主要作用,但我们的发现强调了阿克拉霉素在细胞环境中对 TOP2B 的先前未预料到的影响。