Larsen A K, Gobert C
Institut Gustave-Roussy, Laboratory of Biology and Pharmacology of DNA Topoisomerases 39, Rue Camille Desmoulins, Villejuif, 94805, France.
Pathol Oncol Res. 1999;5(3):171-8. doi: 10.1053/paor.1999.0209.
Mammalian DNA topoisomerase I is a multifunctional enzyme which is essential for embryonal development. In addition to its classical DNA nicking-closing activities which are needed for relaxation of supercoiled DNA, topoisomerase I can phosphorylate certain splicing factors. The enzyme is also involved in transcriptional regulation through its ability to associate with other proteins in the TFIID-, and possibly TFIIH-, transcription complexes, and is implicated in the recognition of DNA lesions. Finally, topoisomerase I is a recombinase which can mediate illegitimate recombination. A crucial reaction intermediate during relaxation of DNA is the formation of a DNA-topoisomerase I complex (the cleavable complex) where topoisomerase I is covalently linked to a 3 -end of DNA thereby creating a single stranded DNA break. Cleavable complexes are also formed in the vicinity of DNA lesions and in the presence of the antitumor agent, camptothecin. While formation of cleavable complexes may be necessary for the initial stages of the DNA damage response, these complexes are also potentially dangerous to the cell due to their ability to mediate illegitimate recombination, which can lead to genomic instability and oncogenesis. Thus the levels and stability of these complexes have to be strictly regulated. This is obtained by maintaining the enzyme levels relatively constant, by limiting the stability of the cleavable complexes through physical interaction with the oncogene suppressor protein p53 and by degradation of the topoisomerase I by the proteasome system. Emerging evidence suggest that these regulatory functions are perturbed in tumor cells, explaining at the same time why topoisomerase I activities so often are increased in certain human tumors, and why these cells are sensitized to the cytotoxic effects of camptothecins.
哺乳动物DNA拓扑异构酶I是一种多功能酶,对胚胎发育至关重要。除了其经典的DNA切口-封闭活性(这是松弛超螺旋DNA所必需的)外,拓扑异构酶I还能使某些剪接因子磷酸化。该酶还通过与TFIID转录复合物以及可能的TFIIH转录复合物中的其他蛋白质结合的能力参与转录调控,并与DNA损伤的识别有关。最后,拓扑异构酶I是一种重组酶,可介导非法重组。DNA松弛过程中的一个关键反应中间体是DNA-拓扑异构酶I复合物(可切割复合物)的形成,其中拓扑异构酶I与DNA的3'-末端共价连接,从而产生单链DNA断裂。在DNA损伤附近以及存在抗肿瘤药物喜树碱的情况下也会形成可切割复合物。虽然可切割复合物的形成可能是DNA损伤反应初始阶段所必需的,但由于它们介导非法重组的能力,这些复合物对细胞也有潜在危险,这可能导致基因组不稳定和肿瘤发生。因此,必须严格调节这些复合物的水平和稳定性。这是通过保持酶水平相对恒定、通过与抑癌蛋白p53的物理相互作用限制可切割复合物的稳定性以及通过蛋白酶体系统降解拓扑异构酶I来实现的。新出现的证据表明,这些调节功能在肿瘤细胞中受到干扰,这同时解释了为什么拓扑异构酶I的活性在某些人类肿瘤中经常增加,以及为什么这些细胞对喜树碱的细胞毒性作用敏感。