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Influence of the 5'-terminal sequences on the 5'-UTR structure of HIV-1 genomic RNA.

ABSTRACT: The 5'-UTR of HIV-1 genomic RNA is known to form specific structures and has important functions. There are three 5'-terminal sequences, G1, G2 and G3, with different localizations in the cell and virion particles as well as different efficiencies in translation and reverse transcription reactions. In the present study, the structural characteristics of the joint region between the TAR and PolyA stems was analysed, and it was found that small differences in the 5'-terminus affect the conformational characteristics of the stem-loop structures. In the G1 form, the two stems form a coaxial stem, whereas in the G2 and G3 forms, the two stems are structurally independent of each other. In the case of the G1 form, the 3'-flanking nucleotides of the PolyA stem are included in the stable coaxial stem structure, which may affect the rest of the 5'-UTR structure. This result demonstrates that the local conformation of this functionally key region has an important role in the function of the 5'-UTR.

SUBMITTER: Obayashi CM

PROVIDER: S-EPMC8149415 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature

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Influence of the 5'-terminal sequences on the 5'-UTR structure of HIV-1 genomic RNA.

Obayashi Camille Michiko CM Shinohara Yoko Y Masuda Takao T Kawai Gota G

Scientific reports 20210525 1

The 5'-UTR of HIV-1 genomic RNA is known to form specific structures and has important functions. There are three 5'-terminal sequences, G1, G2 and G3, with different localizations in the cell and virion particles as well as different efficiencies in translation and reverse transcription reactions. In the present study, the structural characteristics of the joint region between the TAR and PolyA stems was analysed, and it was found that small differences in the 5'-terminus affect the conformatio ...[more]

PMID: 34035384

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Project description:BackgroundDuring HIV-1 maturation, Gag and Gag-Pol polyproteins are proteolytically cleaved and the capsid protein polymerizes to form the honeycomb capsid lattice. HIV-1 integrase (IN) binds the viral genomic RNA (gRNA) and impairment of IN-gRNA binding leads to mis-localization of the nucleocapsid protein (NC)-condensed viral ribonucleoprotein complex outside the capsid core. IN and NC were previously demonstrated to bind to the gRNA in an orthogonal manner in virio; however, the effect of IN binding alone or simultaneous binding of both proteins on gRNA structure is not yet well understood.ResultsUsing crosslinking-coupled selective 2'-hydroxyl acylation analyzed by primer extension (XL-SHAPE), we characterized the interaction of IN and NC with the HIV-1 gRNA 5'-untranslated region (5'-UTR). NC preferentially bound to the packaging signal (Psi) and a UG-rich region in U5, irrespective of the presence of IN. IN alone also bound to Psi but pre-incubation with NC largely abolished this interaction. In contrast, IN specifically bound to and affected the nucleotide (nt) dynamics of the apical loop of the transactivation response element (TAR) and the polyA hairpin even in the presence of NC. SHAPE probing of the 5'-UTR RNA in virions produced from allosteric IN inhibitor (ALLINI)-treated cells revealed that while the global secondary structure of the 5'-UTR remained unaltered, the inhibitor treatment induced local reactivity differences, including changes in the apical loop of TAR that are consistent with the in vitro results.ConclusionsOverall, the binding interactions of NC and IN with the 5'-UTR are largely orthogonal in vitro. This study, together with previous probing experiments, suggests that IN and NC binding in vitro and in virio lead to only local structural changes in the regions of the 5'-UTR probed here. Accordingly, disruption of IN-gRNA binding by ALLINI treatment results in local rather than global secondary structure changes of the 5'-UTR in eccentric virus particles.

Dataset Information

Influence of the 5'-terminal sequences on the 5'-UTR structure of HIV-1 genomic RNA.

Publications

Influence of the 5'-terminal sequences on the 5'-UTR structure of HIV-1 genomic RNA.

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