Hoeffler W K, Roeder R G
Cell. 1985 Jul;41(3):955-63. doi: 10.1016/s0092-8674(85)80076-3.
Nuclear extracts from adenovirus-infected HeLa cells harvested early in infection demonstrated a markedly increased capacity for transcription by RNA polymerase III of exogenous VA RNA genes, as well as cloned tRNA and 5S RNA genes. In contrast, no enhanced transcription was observed in extracts from cells infected with an E1A deletion mutant. Moreover, cells co-transfected with the VA- and E1A-containing plasmids showed markedly higher levels of VA RNA synthesis than did cells transfected with the VA-containing plasmid alone. Although analysis of high ionic strength extracts revealed that the enhancement of pol III transcription persists late in infection, moderate ionic strength extracts indicated that transcription factor IIIC becomes limiting. Chromatographic fractionation and complementation analysis of extracts from mock- and virus-infected cells indicated that the factor(s) responsible for the enhanced activity was localized entirely in the fraction containing transcription factor IIIC.
感染早期收获的腺病毒感染的HeLa细胞的核提取物显示,其对外源VA RNA基因以及克隆的tRNA和5S RNA基因进行RNA聚合酶III转录的能力显著增强。相比之下,感染E1A缺失突变体的细胞提取物中未观察到转录增强。此外,与单独转染含VA质粒的细胞相比,共转染含VA和E1A质粒的细胞显示出明显更高水平的VA RNA合成。尽管对高离子强度提取物的分析表明,感染后期pol III转录的增强持续存在,但中等离子强度提取物表明转录因子IIIC成为限制因素。对模拟感染和病毒感染细胞提取物的色谱分级分离和互补分析表明,负责增强活性的因子完全定位于含有转录因子IIIC的级分中。