Le J Y, Defendi V
Department of Pathology, New York University Medical Center, New York 10016.
J Virol. 1988 Nov;62(11):4420-6. doi: 10.1128/JVI.62.11.4420-4426.1988.
A 4.4-kilobase DNA fragment (T4.4) from a human tumor (comprising part of the human papillomavirus type 16 E6 promoter; the E6, E7, and part of the E1 open reading frames; and cellular sequences) was found to be competent to fully transform NIH 3T3 cells. This competency resides in the whole hybrid DNA fragment, since the separate viral or cellular DNA sequences were not active. Abundant E6-E7 transcripts were found in the transformed cells. When the cellular fragments were substituted with polyadenylation sequences from polyomavirus or simian virus 40 DNA, little or no restoration of transforming activity was observed. In experiments in which an exogenous reporting gene, that for chloramphenicol acetyltransferase, was used, the possibility was excluded that the cellular flanking sequences act as a traditional enhancer; yet, when the cellular sequences were placed downstream of a chloramphenicol acetyltransferase expression vector (pSV2 CAT), activity of the reference gene was clearly enhanced. These results indicate that DNA containing human papillomavirus type 16 open reading frames E6 and E7 isolated from the genome of a human tumor has transforming potential, that this potential is realized when the viral DNA is joined to cellular sequences, and that the cellular sequences function in a more complex way than by simply providing polyadenylation signals.
从一个人类肿瘤中分离出一个4.4千碱基的DNA片段（T4.4）（包含人乳头瘤病毒16型E6启动子的一部分、E6、E7以及E1开放阅读框的一部分和细胞序列），发现其能够完全转化NIH 3T3细胞。这种转化能力存在于整个杂交DNA片段中，因为单独的病毒或细胞DNA序列没有活性。在转化细胞中发现了大量的E6 - E7转录本。当用多瘤病毒或猿猴病毒40 DNA的聚腺苷酸化序列替代细胞片段时，几乎没有观察到转化活性的恢复。在使用外源性报告基因氯霉素乙酰转移酶基因的实验中，排除了细胞侧翼序列作为传统增强子的可能性；然而，当将细胞序列置于氯霉素乙酰转移酶表达载体（pSV2 CAT）的下游时，报告基因的活性明显增强。这些结果表明，从人类肿瘤基因组中分离出的含有人类乳头瘤病毒16型开放阅读框E6和E7的DNA具有转化潜力，当病毒DNA与细胞序列连接时这种潜力得以实现，并且细胞序列的功能比仅仅提供聚腺苷酸化信号更为复杂。
