CONTROL OF HUMAN PAPILLOMAVIRUS TYPE-11 ORIGIN OF REPLICATION BY THE E2 FAMILY OF TRANSCRIPTION REGULATORY PROTEINS
CONTROL OF HUMAN PAPILLOMAVIRUS TYPE-11 ORIGIN OF REPLICATION BY THE E2 FAMILY OF TRANSCRIPTION REGULATORY PROTEINS
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DOI：
10.1128/jvi.66.9.5224-5231.1992
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发表时间：
1992-09-01
影响因子：
5.4
通讯作者：
CHOW, LT
中科院分区：
文献类型：
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作者：
CHIANG, CM;DONG, G;CHOW, LT
人乳头瘤病毒11型（HPV-11）DNA的复制需要全长病毒E1和E2蛋白（C.- M. Chiang，M. Ustav，A. Stenlund，T. F.霍，T. R. Broker和L. T. Chow，Proc. Natl. Acad. Sci. USA 89：5799-5803，1992）。使用瞬时转染的亚基因组HPV DNA到仓鼠CHO和人类293细胞，我们已经定位的复制起点（ori）的上游调控区跨越核苷酸1的80 bp的片段。它与E6启动子区重叠，并含有一个短的富含A+ T的片段和一个与牛乳头瘤病毒1型（BPV-1）E1蛋白在BPV-1 ori中的结合位点同源的序列。然而，与BPV-1 ori不同，对于BPV-1 ori，半个E2-应答序列（E2-RS）或结合位点就足够了，HPV-11 ori需要完整的结合位点。当存在额外的E2-RS时，复制更有效。当提供E1和E2蛋白时，完整的HPV-11基因组也在两种细胞系中复制。缺乏E2转录反式激活所必需的N-末端结构域的转录抑制蛋白的表达载体不支持与E1表达载体协作的复制。相反，与阻遏物表达载体的共转染抑制ori复制的E1和E2蛋白。这些结果证明了E2蛋白的N-末端结构域在DNA复制中的重要性，并表明E2蛋白家族正向和负向调节病毒DNA复制和E6启动子转录。
Replication of human papillomavirus type 11 (HPV-11) DNA requires the full-length viral E1 and E2 proteins (C.-M. Chiang, M. Ustav, A. Stenlund, T. F. Ho, T. R. Broker, and L. T. Chow, Proc. Natl. Acad. Sci. USA 89:5799-5803, 1992). Using transient transfection of subgenomic HPV DNA into hamster CHO and human 293 cells, we have localized an origin of replication (ori) to an 80-bp segment in the upstream regulatory region spanning nucleotide 1. It overlaps the E6 promoter region and contains a short A+T-rich segment and a sequence which is homologous to the binding site of the bovine papillomavirus type 1 (BPV-1) E1 protein in the BPV-1 ori. However, unlike the BPV-1 ori, for which half an E2-responsive sequence (E2-RS) or binding site suffices, an intact binding site is essential for the HPV-11 ori. Replication was more efficient when additional E2-RSs were present. The intact HPV-11 genome also replicated in both cell lines when supplied with E1 and E2 proteins. Expression vectors of transcription repressor proteins that lack the N-terminal domain essential for E2 transcriptional trans activation did not support replication in collaboration with the E1 expression vector. Rather, cotransfection with the repressor expression vectors inhibited ori replication by the E1 and E2 proteins. These results demonstrate the importance of the N-terminal domain of the E2 protein in DNA replication and indicate that the family of E2 proteins positively and negatively regulates both viral DNA replication and E6 promoter transcription.