p53-independent and -dependent requirements for E1B-55K in adenovirus type 5 replication
p53-independent and -dependent requirements for E1B-55K in adenovirus type 5 replication
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DOI：
10.1128/jvi.73.7.5333-5344.1999
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发表时间：
1999-07-01
影响因子：
5.4
通讯作者：
Berk, AJ
中科院分区：
文献类型：
--
作者：
Harada, JN;Berk, AJ
腺病毒 5 型突变体 dl1520 先前被设计为 E1B-55K 功能完全缺陷。最近，这种突变体(也称为ONYX-015)被认为优先在p53(-)和一些p53(+)肿瘤细胞系中复制，但在原代培养细胞中减弱(C.Heise、A.Sampson-Johannes、A.Williams、F.McCormick、D.D.F.Hoff和D.H.Firn，Nat.Med.3：639-645， 1997）。有人认为dl1520可能被用作“灵丹妙药”，可以选择性地裂解肿瘤细胞而不伤害正常组织。然而，我们在此报告，dl1520复制独立于p53基因型，并且在一些原代培养的人类细胞中有效发生，表明突变病毒不具有肿瘤选择性。虽然它不是唯一的宿主范围决定因素，但当在表达温度敏感 p53 (H1299-tsp53) 的细胞系中进行分析时，p53 功能确实减少了 dl1520 复制 (K. L. Fries, W E. Miller, and N. Raab-Traub, J. Virol. 70:8653-8659, 1996)。正如早期在 HeLa 细胞中发现的其他 E1B-55K 突变体（Y. Ho、R. Gales 和 J. Williams，Virology 122：109-124，1982），dl1520 复制在 H1299 细胞中是温度依赖性的。当 H1299-tsp53 细胞中 p53 功能在低温下恢复时，它对病毒 DNA 复制和晚期病毒细胞质 mRNA 的积累造成了适度的缺陷。然而，在 H1299 和 H1299-tsp53 细胞中，晚期病毒蛋白质合成的缺陷似乎比晚期病毒 nnRNA 水平的适度缺陷所能解释的要大得多。因此，我们提出，除了对抗 p53 功能和调节病毒和细胞 mRNA 核转运外，E1B-55K 还刺激晚期病毒 mRNA 翻译。
The adenovirus type 5 mutant dl1520 was engineered previously to be completely defective for E1B-55K functions. Recently, this mutant (also known as ONYX-015) has been suggested to replicate preferentially in p53(-) and some p53(+) tumor cell lines but to be attenuated in primary cultured cells (C. Heise, A. Sampson-Johannes, A. Williams, F. McCormick, D. D. F. Hoff, and D. H. Firn, Nat. Med. 3:639-645, 1997). It has been suggested that dl1520 might be used as a "magic bullet" that could selectively lyse tumor cells without harm to normal tissues. However, we report here that dl1520 replication is independent of p53 genotype and occurs efficiently in some primary cultured human cells, indicating that the mutant virus does not possess a tumor selectivity. Although it was not the sole host range determinant, p53 function did reduce dl1520 replication when analyzed in a cell line expressing temperature-sensitive p53 (H1299-tsp53) (K. L. Fries, W E. Miller, and N. Raab-Traub, J. Virol. 70:8653-8659, 1996). As found earlier far other E1B-55K mutants in HeLa cells (Y. Ho, R. Gales, and J. Williams, Virology 122:109-124, 1982), dl1520 replication was temperature dependent in H1299 cells. When p53 function was restored at low temperature in H1299-tsp53 cells, it imposed a modest defect in viral DNA replication and accumulation of late viral cytoplasmic mRNA. However, in both H1299 and H1299-tsp53 cells, the defect in late viral protein synthesis appeared to be much greater than could be accounted for by the modest defects in late viral nnRNA levels. We therefore propose that in addition to countering p53 function and modulating viral and cellular mRNA nuclear transport, E1B-55K also stimulates late viral mRNA translation.