Rohlfs M, Winkenbach S, Meyer S, Rupp T, Dürst M
Institut für Virusforschung, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Virology. 1991 Jul;183(1):331-42. doi: 10.1016/0042-6822(91)90146-3.
Human papillomavirus type-16 (HPV-16) transcription in two human keratinocyte cell lines (HPK) immortalized by transfection of viral DNA in vitro was analyzed by nucleotide sequencing of cDNA clones, and in addition by primer extension analysis and S1 nuclease and exonuclease VII digestion of poly(A)+ RNA. A novel mRNA species which probably initiates in the E7 ORF and in which the 5'-part of the E1 ORF (splice donor at position (pos.) 880) is joined to an exon comprising the entire E2 ORF (splice acceptor at pos. 2708) was found in both cell lines. This mRNA has the potential to encode a full-length E2 protein, which is known to function as a repressor of transcription initiated at P97. cDNAs derived from the late region of the viral genome and the use of a late polyadenylation signal at pos. 7320-7325 are described. In agreement with RNA data published by others the major promoter for HPV-16 transcription is located at pos. 97. mRNA species encoding full-length or truncated forms of the E6 protein, and species characterized by an E1i [symbol see text] E4 splice junction (which provided the E4 open reading frame (ORF) with an ATG triplet) were identified.
通过对cDNA克隆进行核苷酸测序，以及对多聚腺苷酸加尾RNA进行引物延伸分析、S1核酸酶和核酸外切酶VII消化，对体外转染病毒DNA永生化的两个人类角质形成细胞系（HPK）中的人乳头瘤病毒16型（HPV - 16）转录进行了分析。在这两个细胞系中均发现了一种新的mRNA种类，它可能起始于E7开放阅读框（ORF），其中E1 ORF的5'部分（第880位的剪接供体）与一个包含整个E2 ORF的外显子相连（第2708位的剪接受体）。这种mRNA有潜力编码全长E2蛋白，已知该蛋白可作为在P97起始的转录的抑制因子。描述了源自病毒基因组晚期区域的cDNA以及在第7320 - 7325位使用晚期多聚腺苷酸化信号的情况。与其他人发表的RNA数据一致，HPV - 16转录的主要启动子位于第97位。鉴定出了编码E6蛋白全长或截短形式的mRNA种类，以及以E1i [符号见原文] E4剪接连接为特征的种类（该剪接连接为E4开放阅读框（ORF）提供了一个ATG三联体）。
