RNA replication for the paramyxovirus simian virus 5 requires an internal repeated (CGNNNN) sequence motif
RNA replication for the paramyxovirus simian virus 5 requires an internal repeated (CGNNNN) sequence motif
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DOI：
10.1128/jvi.73.1.805-809.1999
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发表时间：
1999-01-01
影响因子：
5.4
通讯作者：
Parks, GD
中科院分区：
文献类型：
--
作者：
Murphy, SK;Parks, GD
副粘病毒猿病毒 5 (SV5) 的功能性 RNA 复制启动子需要两个必需且不连续的元件：3' 末端的 19 个碱基（保守区 I）和包含在 L 蛋白基因编码区内的 18 个碱基内部区域（保守区 II [CRII]）。使用反向遗传学系统来确定内部 CRII 元件在 RNA 复制中发挥作用的序列要求。构建了一系列回录缺陷干扰（DI）RNA类似物，使其在组成CRII的18个核苷酸中含有点突变，并分析了它们的相对复制水平。结果表明，SV5 DI RNA复制通过替换两个CG二核苷酸而减少，这两个CG二核苷酸在核衣壳模板中位于CRII核苷酸的前两个六聚体的前两个位置。 CRII 内其他碱基的取代不会减少 RNA 合成，因此，两个连续的 5'-CGNNNN-3' 六聚体形成 SV5 CRII 启动子元件中的重要序列。 SV5 前导序列和抗拖尾启动子内的 CG 二核苷酸位置在风疹病毒属的其他成员中高度保守，但该基序在序列和位置上都与先前为仙台病毒鉴定的基序显着不同。讨论了 CRII 内部启动子元件在副粘病毒 RNA 复制中的可能作用。
A functional RNA replication promoter for the paramyxovirus simian virus 5 (SV5) requires two essential and discontinuous elements: 19 bases at the 3' terminus (conserved region I) and an 18-base internal region (conserved region II [CRII]) that is contained within the coding region of the L protein gene. A reverse-genetics system was used to determine the sequence requirements for the internal CRII element to function in RNA replication. A series of copyback defective interfering (DI) RNA analogs were constructed to contain point mutations in the 18 nucleotides composing CRII, and their relative replication levels were analyzed, The results indicated that SV5 DI RNA replication was reduced by substitutions for two CG dinucleotides, which in the nucleocapsid template are in the first two positions of the first two hexamers of CRII nucleotides. Substitutions for other bases within CRII did not reduce RNA synthesis, Thus, two consecutive 5'-CGNNNN-3' hexamers form an important sequence in the SV5 CRII promoter element. The position of the CG dinucleotide within the SV5 leader and antitrailer promoters was highly conserved among other members of the Rubulavirus genus, but this motif differed significantly in both sequence and position from that previously identified for Sendai virus. The possible roles of the CRII internal promoter element in paramyxovirus RNA replication are discussed.