HIV-1-encoded antisense RNA suppresses viral replication for a prolonged period.
HIV-1-encoded antisense RNA suppresses viral replication for a prolonged period.
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DOI：
10.1186/1742-4690-9-38
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发表时间：
2012-05-08
期刊：
影响因子：
3.3
通讯作者：
Watanabe T
中科院分区：
文献类型：
--
作者：
Kobayashi-Ishihara M;Yamagishi M;Hara T;Matsuda Y;Takahashi R;Miyake A;Nakano K;Yamochi T;Ishida T;Watanabe T
最近的证据提出了一个新的概念，哺乳动物天然反义RNA通过调节几个基因的表达在细胞内稳态中发挥重要作用。逆转录病毒反义RNA的鉴定和表征将为复制和发病机制提供新的见解。HIV-1编码的反义RNA已被报道，但其结构和功能仍有待研究。我们试图鉴定和表征HIV-1的反义RNA及其在病毒感染中的功能。HEK 293 T细胞的转录物的表征表明，可以表达各种反义转录物，所述HEK 293 T细胞用具有HIV-1 NL 4 -3 DNA的表达质粒以反义方向瞬时转染。通过筛选和表征HIV-1 NL 4 -3感染细胞中的反义RNA，我们定义了HIV-1反义RNA的主要形式的一级结构，其对应于先前报道的ASP mRNA的变体。在急性或慢性感染的细胞系和急性感染的人外周血单核细胞中，该2.6 kb RNA从3′ LTR的U3区转录并终止于env区。报告基因分析清楚地表明，HIV-1 LTR在相反方向上具有启动子活性。突变分析表明NF-κ B结合位点参与反义转录的调节。反义RNA定位于感染细胞的细胞核中。这种反义RNA的表达抑制HIV-1复制超过一个月。此外，这种反义RNA的特异性敲低增强了HIV-1基因的表达和复制。本研究的结果确定了从HIV-1 NL 4 -3前病毒表达的反义RNA的主要形式的准确结构，并证明了其核定位。功能研究共同证明了反义RNA在病毒复制中的新作用。因此，我们提出了一种新的病毒机制，自我限制HIV-1复制，并提供了新的见解病毒的生命周期。
Recent evidence proposes a novel concept that mammalian natural antisense RNAs play important roles in cellular homeostasis by regulating the expression of several genes. Identification and characterization of retroviral antisense RNA would provide new insights into mechanisms of replication and pathogenesis. HIV-1 encoded-antisense RNAs have been reported, although their structures and functions remain to be studied. We have tried to identify and characterize antisense RNAs of HIV-1 and their function in viral infection. Characterization of transcripts of HEK293T cells that were transiently transfected with an expression plasmid with HIV-1NL4–3 DNA in the antisense orientation showed that various antisense transcripts can be expressed. By screening and characterizing antisense RNAs in HIV-1NL4–3-infected cells, we defined the primary structure of a major form of HIV-1 antisense RNAs, which corresponds to a variant of previously reported ASP mRNA. This 2.6 kb RNA was transcribed from the U3 region of the 3′ LTR and terminated at the env region in acutely or chronically infected cell lines and acutely infected human peripheral blood mononuclear cells. Reporter assays clearly demonstrated that the HIV-1 LTR harbours promoter activity in the reverse orientation. Mutation analyses suggested the involvement of NF-κΒ binding sites in the regulation of antisense transcription. The antisense RNA was localized in the nuclei of the infected cells. The expression of this antisense RNA suppressed HIV-1 replication for more than one month. Furthermore, the specific knockdown of this antisense RNA enhanced HIV-1 gene expression and replication. The results of the present study identified an accurate structure of the major form of antisense RNAs expressed from the HIV-1NL4–3 provirus and demonstrated its nuclear localization. Functional studies collectively demonstrated a new role of the antisense RNA in viral replication. Thus, we suggest a novel viral mechanism that self-limits HIV-1 replication and provides new insight into the viral life cycle.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.3
作者：
Landry S;Halin M;Lefort S;Audet B;Vaquero C;Mesnard JM;Barbeau B
通讯作者：
Barbeau B
影响因子：
7.7
作者：
Bell, SD;Brinkman, AB;Jackson, SP
通讯作者：
Jackson, SP
DOI：
10.1073/pnas.86.7.2365
发表时间：
1989-04-01
影响因子：
11.1
作者：
FOLKS, TM;CLOUSE, KA;FAUCI, AS
通讯作者：
FAUCI, AS
影响因子：
5.3
作者：
Berteaux, Nathalie;Aptel, Nathalie;Adriaenssens, Eric
通讯作者：
Adriaenssens, Eric