Precise mapping of the replication and transcription promoters of human parainfluenza virus type 3
Precise mapping of the replication and transcription promoters of human parainfluenza virus type 3
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DOI：
10.1006/viro.2000.0223
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发表时间：
2000-03-30
期刊：
影响因子：
3.7
通讯作者：
Banerjee, AK
中科院分区：
文献类型：
--
作者：
Hoffman, MA;Banerjee, AK
已知副粘病毒和弹状病毒的基因组和反基因组RNA的末端RNA区域含有指导RNA复制和转录所必需的序列。弹状病毒和副粘病毒的负义基因组RNA的3 '末端（前导区）被称为前导（Le）启动子，并指导正义复制和转录产物的合成。反基因组的3 '末端被称为尾部互补（TrC）启动子并指导基因组RNA的合成。通过在HPIV3微复制子的相应区域中产生突变，其中病毒蛋白编码序列被荧光素酶基因取代，我们能够精确地定义参与指导HPIV3正链复制的前导启动子的元件。核苷酸1至12（从末端）形成了复制的关键域。从核苷酸13到55的区域对于复制很重要，但不是至关重要的，而位置79、85和91处的G残基构成了另一个对于复制至关重要的结构域。还显示TrC启动子与Le启动子相似，但不相同。TrC启动子的核苷酸1至12对于基因组RNA的合成是关键的，尽管特定位置与Le启动子的相应位置表现不同。虽然这些突变中的许多不能被分析用于转录，因为它们完全废除了基因组RNA合成（转录模板），但我们惊讶地发现，前导启动子中没有减少复制的突变对转录有任何显著影响。然而，在前导序列之后和荧光素酶信息之前的基因间序列和基因起始信号中的突变严重降低了转录，但不是复制。(C)北京大学出版社.
The terminal RNA regions of the genomic and antigenomic RNAs of the paramyxoviruses and rhabdoviruses are known to contain sequences essential for directing RNA replication and transcription. The 3' terminus (leader region) of the negative-sense, genomic RNA of the rhabdoviruses and paramyxoviruses is known as the leader (Le) promoter and directs synthesis of positive-sense replication and transcription products. The 3' terminus of the antigenome is termed the trailer complementary (TrC) promoter and directs the synthesis of genomic RNA. By creating mutations in the corresponding regions of an HPIV3 minireplicon in which the viral protein coding sequences were replaced by the luciferase gene, we were able to precisely define the elements of the leader promoter involved in directing positive-strand replication of HPIV3. Nucleotides 1 through 12 (from the terminus) formed a domain critical for replication. The region from nucleotides 13 through 55 was important but not crucial for replication, while G residues at positions 79, 85, and 91 comprised another domain critical for replication. it was also shown that the TrC promoter is similar, though not identical, to the Le promoter. Nucleotides 1 through 12 of the TrC promoter were critical for synthesis of genomic RNA, though specific positions behaved differently from the corresponding positions of the Le promoter. While many of these mutations could not be analyzed for transcription because they completely abrogated genomic RNA synthesis (the template for transcription), we were surprised to find that no mutations in the leader promoter which decreased replication had any significant effect on transcription. However, mutations in the intergenic sequence and gene start signal following the leader and preceding the luciferase message severely decreased transcription, but not replication. (C) 2000 Academic Press.