DNA聚合酶参与培养的中国仓鼠细胞中苯并[a]芘DNA损伤的修复

Fuminori Otsuka, Takafumi Ochi, Motoyasu Ohsawa
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引用次数: 6

摘要

采用碱性蔗糖梯度沉降法研究了s9激活的苯并[a]芘(B[a]P)诱导DNA单链断裂及其在培养的中国仓鼠V79细胞中的修复机制。5 μg/ml的B[a]P作用于V79细胞3 h后,观察到DNA单链断裂的明显诱导。这些DNA损伤在去除B[a]P后4 h内修复至对照水平。同时加入DNA修复合成抑制剂,1-β- d -阿拉伯糖醛酸胞嘧啶(araC)和羟基脲与B[a]P并没有增加DNA单链断裂的形成。然而,当这些抑制剂在去除B[a]P后加入时,它们显著阻断了dna链断裂的重新连接。另一方面,当DNA聚合酶α特异性抑制剂aphidicolin代替araC时,观察到重新连接的部分抑制,并且进一步添加DNA聚合酶β抑制剂2 ',3 ' -二脱氧胸腺嘧啶增强了抑制作用。这些结果表明,B[a] p诱导的V79细胞DNA单链断裂主要可以通过DNA聚合酶α和非α聚合酶(推测为聚合酶β)的切除修复来修复。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Involvement of DNA polymerases in the repair of DNA damage by benzo[a]pyrene in cultured Chinese hamster cells

Mechanisms for induction of single-strand scissions in DNA by S9-activated benzo[a]pyrene (B[a]P) and their repair in cultured Chinese hamster V79 cells were investigated with inhibitors of DNA-repair synthesis using alkaline sucrose gradient sedimentation analysis.

The marked induction of single-strand scissions in DNA was observed following 3 h treatment of V79 cells with 5 μg/ml of B[a]P. These DNA lesions were repaired to the control level within 4 h after removal of B[a]P. The simultaneous addition of inhibitors of DNA-repair synthesis, 1-β-D-arabinofuranosylcytosine (araC) plus hydroxyurea with B[a]P did not increase the formation of DNA single-strand scissions. When these inhibitors were added after removal of B[a]P, however, they significantly blocked the rejoining of DNA-strand scissions. On the other hand, when aphidicolin, a specific inhibitor of DNA polymerase α, was used instead of araC, a partial inhibition of the rejoining was observed, and further addition of 2′,3′-dideoxythymidine, an inhibitor of DNA polymerase β, augmented the inhibitory effect. These results indicate that B[a]P-induced single-strand scissions of DNA in V79 cells could be repaired mostly by excision repair which involved DNA polymerase α and a non-α polymerase, presumably polymerase β.

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