Characterization of the transcription profile of adeno-associated virus type 5 reveals a number of unique features compared to previously characterized adeno-associated viruses
Characterization of the transcription profile of adeno-associated virus type 5 reveals a number of unique features compared to previously characterized adeno-associated viruses
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DOI：
10.1128/jvi.76.24.12435-12447.2002
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发表时间：
2002-12-01
影响因子：
5.4
通讯作者：
Pintel, DJ
中科院分区：
文献类型：
--
作者：
Qiu, JM;Nayak, R;Pintel, DJ
我们报道了病毒感染后产生的5型腺相关病毒(AAV5)RNA的初步特征以及AAV5复制感染性克隆的构建。AAV5的基本转录图谱与AAV2相似，但也存在显著差异。对AAV5转录本的定位表明，在AAV5反向末端重复序列(ITR)中有一个有效的转录起始点，而对AAV5内含子的定位表明，它比AAV2的转录起始点小得多。此外，与AAV2的情况相反，在将缺失ITRS的AAV5rep/Cap克隆导入293细胞后，既不需要Rep蛋白也不需要额外的腺病毒基因产物来实现有效的启动子活性和前mRNA剪接。也许最令人惊讶的是，从AAV5 P7和P19启动子产生的RNA在基因组中心的内含子区域内的位置被高效地多腺化。由于P7和P19产生的转录本在这个位点上被多聚腺苷化，而不是剪接，所以Rep78和Rep52是AAV5感染过程中仅检测到的Rep蛋白。
We report the initial characterization of adeno-associated virus type 5 (AAV5) RNAs generated following viral infection and the construction of a replicating infectious clone of AAV5. While the basic transcription profile of AAV5 was similar to that of AAV2, there were also significant differences. Mapping of the AAV5 transcripts demonstrated an efficient transcription initiation site within the AAV5 inverted terminal repeat (ITR), and mapping of the AAV5 intron revealed that it is considerably smaller than that of AAV2. Furthermore, in contrast to the case for AAV2, neither the Rep protein nor additional adenovirus gene products were required to achieve efficient promoter activity and pre-mRNA splicing following transfection of an AAV5 rep/cap plasmid clone lacking the ITRs into 293 cells. Perhaps most surprisingly, RNAs generated from both the AAV5 P7 and P19 promoters were efficiently polyadenylated at a site lying within the intronic region in the center of the genome. Because P7- and P19-generated transcripts are polyadenylated at this site and not spliced, Rep78 and Rep52 were the only Rep proteins detected during AAV5 infection.