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首页> 外文期刊>Cell Research >Expression of DNA-dependent protein kinase in human granulocytes.
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Expression of DNA-dependent protein kinase in human granulocytes.

机译：DNA依赖性蛋白激酶在人粒细胞中的表达。

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摘要

Human polymorphonuclear leukocytes (PMN) have been reported to completely lack of DNA-dependent protein kinase (DNA-PK) which is composed of Ku protein and the catalytic subunit DNA-PKcs, needed for nonhomologous end-joining (NHEJ) of DNA double-strand breaks. Promyelocytic HL-60 cells express a variant form of Ku resulting in enhanced radiation sensitivity. This raises the question if low efficiency of NHEJ, instrumental for the cellular repair of oxidative damage, is a normal characteristic of myeloid differentiation. Here we confirmed the complete lack of DNA-PK in PMN protein extracts, and the expression of the truncated Ku86 variant form in HL-60. However, this degradation of DNA-PK was shown to be due to a DNA-PK-degrading protease in PMN and HL-60. In addition, by using a protease-resistant whole cell assay, both Ku86 and DNA-PKcs could be demonstrated in PMN, suggesting the previously reported absence in PMN of DNA-PK to be an artefact. The levels of Ku86 and DNA-PKcs were much reduced in PMN, as compared with that of the lymphocytes, whereas HL-60 displayed a markedly elevated DNA-PK concentration. In conclusion, our findings provide evidence of reduced, not depleted expression of DNA-PK during the mature stages of myeloid differentiation.
机译：据报道，人类多形核白细胞（PMN）完全缺乏DNA依赖性蛋白激酶（DNA-PK），该蛋白激酶由Ku蛋白和催化亚基DNA-PKcs组成，是DNA双链的非同源末端连接（NHEJ）所需的。链断裂。早幼粒细胞HL-60细胞表达Ku的变体形式，导致增强的辐射敏感性。这就提出了一个问题，即NHEJ的低效率（有助于细胞氧化损伤的修复）是否是骨髓分化的正常特征。在这里，我们证实了PMN蛋白提取物中完全缺乏DNA-PK，并且在HL-60中表达了截短的Ku86变体形式。但是，这种DNA-PK的降解显示是由于PMN和HL-60中的DNA-PK降解蛋白酶引起的。另外，通过使用耐蛋白酶的全细胞测定法，可以在PMN中证明Ku86和DNA-PKcs，这表明先前报道的在PMN中不存在DNA-PK是伪像。与淋巴细胞相比，PMN中Ku86和DNA-PKcs的水平大大降低，而HL-60的DN​​A-PK浓度显着升高。总之，我们的发现提供了在骨髓分化成熟阶段DNA-PK表达减少而不减少的证据。

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