کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5532959 | 1402090 | 2017 | 14 صفحه PDF | دانلود رایگان |
- The RNA sequence necessary and sufficient to mediate HIV-1 genome packaging has not been defined.
- The sequence required to package RNA into HIV-1 particles was determined using two reporter RNAs.
- The HIV-1 5â² UTR and the first 32-nt of gag sequence are not sufficient to mediate packaging.
- Reporter RNAs containing HIV-1 5â² UTR and the 5â² half of gag gene can be packaged into viral particles.
- The role of the gag gene sequence is likely to be indirect but improves the Gag:5â² UTR interaction.
Genome packaging is an essential step to generate infectious HIV-1 virions and is mediated by interactions between the viral protein Gag and cis-acting elements in the full-length RNA. The sequence necessary and sufficient to allow RNA genome packaging into an HIV-1 particle has not been defined. Here, we used two distinct reporter systems to determine the HIV-1 sequence required for heterologous, non-viral RNAs to be packaged into viral particles. Although the 5â² untranslated region (UTR) of the HIV-1 RNA is known to be important for RNA packaging, we found that its ability to mediate packaging relies heavily on the context of the downstream sequences. Insertion of the 5â² UTR and the first 32-nt of gag into two different reporter RNAs is not sufficient to mediate the packaging of these RNA into HIV-1 particles. However, adding the 5â² half of the gag gene to the 5â² UTR strongly facilitates the packaging of two reporter RNAs; such RNAs can be packaged at >Â 50% of the efficiencies of an HIV-1 near full-length vector. To further examine the role of the gag sequence in RNA packaging, we replaced the 5â² gag sequence in the HIV-1 genome with two codon-optimized gag sequences and found that such substitutions only resulted in a moderate decrease of RNA packaging efficiencies. Taken together, these results indicated that both HIV-1 5â² UTR and the 5â² gag sequence are required for efficient packaging of non-viral RNA into HIV-1 particles, although the gag sequence likely plays an indirect role in genome packaging.
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Journal: Journal of Molecular Biology - Volume 429, Issue 16, 4 August 2017, Pages 2542-2555