A STUDY OF THE DIMER FORMATION OF ROUS-SARCOMA VIRUS-RNA AND OF ITS EFFECT ON VIRAL PROTEIN-SYNTHESIS INVITRO
A STUDY OF THE DIMER FORMATION OF ROUS-SARCOMA VIRUS-RNA AND OF ITS EFFECT ON VIRAL PROTEIN-SYNTHESIS INVITRO
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DOI：
10.1093/nar/18.1.119
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发表时间：
1990-01-11
影响因子：
14.9
通讯作者：
DARLIX, JL
中科院分区：
文献类型：
--
作者：
BIETH, E;GABUS, C;DARLIX, JL
到目前为止，所有研究的逆转录病毒的遗传物质都是RNA二聚体，其中两个相同的RNA亚基在其5“末端通过称为二聚体连接结构（DLS）的结构连接。由于DLS的精确位置和结构以及RNA二聚化的机制和作用仍不清楚，我们分析了劳斯肉瘤病毒（RSV）RNA的二聚化过程。为此，我们建立了RSV RNA二聚化的体外模型。利用该模型研究了RSV RNA形成二聚体的过程，并发现RSV的核衣壳蛋白（NCp 12）对二聚体的形成有很强的激活作用。在互补的5“32 P-DNA寡聚体存在下进行RSV RNA二聚化，以探测RSV RNA的单体和二聚体形式。数据表明，RSV RNA的DLS可能定位在从5“端起的位置544-564之间。为了确定RSV RNA二聚化所需的序列，在5“600 nt处产生缺失诱变。结果表明，二聚体启动序列可能位于距离5“端的208-270和400-600位，因此可能包含RSV基因组RNA特异性折叠所需的顺式作用元件。还报道了多蛋白前体Pr 76 gag的合成在RSV RNA二聚化后受到抑制。这些结果表明，二聚体和二聚体化的基因组长度的RSV RNA可能在病毒体形成的过程中，因为它们似乎是在相同的顺式元件，Ea和DLS，和反式作用因子核衣壳蛋白NCp 12的控制下。
The genetic material of all retroviruses examined so far is an RNA dimer where two identical RNA subunits are joined at their 5'' ends by a structure named dimer linkage structure (DLS). Since the precise location and structure of the DLS as well as the mechanism and role(s) of RNA dimerization remain unclear, we analysed the dimerization process of Rous sarcoma virus (RSV) RNA. For this purpose we set up an in vitro model for RSV RNA dimerization. Using this model RSV RNA was shown to form dimeric molecules and this dimerization process was greatly activated by nucleocapsid protein (NCp12) of RSV, Furthermore. RSV RNA dimerization was performed in the presence of complementary 5''32P-DNA oligomers in order to probe the monomer and dimer forms of RSV RNA. Data indicated that the DLS of RSV RNA probably maps between positions 544-564 from the 5'' end. In an attempt to define sequences needed for the dimerization of RSV RNA, deletion mutageneses were generated in the 5'' 600 nt. The results showed that the dimer promoting sequences probably are located within positions 208-270 and 400-600 from the 5''end and hence possibly encompassing the cis-acting elements needed for the specific encapsidation of RSV genomic RNA. Also it is reported that synthesis of the polyprotein precursor Pr76gag is inhibited upon dimerization of RSV RNA. These results suggest that dimerization and encapsidation of genome length RSV RNA might be linked in the course of virion formation since they appear to be under the control of the same cis elements, Eaand DLS, and the trans-acting factor nucleocapsid protein NCp12.