Dataset Information

Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55^Gag and its maturation products.

ABSTRACT: Maturation of the HIV-1 viral particles shortly after budding is required for infectivity. During this process, the Pr55^Gag precursor undergoes a cascade of proteolytic cleavages, and whilst the structural rearrangements of the viral proteins are well understood, the concomitant maturation of the genomic RNA (gRNA) structure is unexplored, despite evidence that it is required for infectivity. To get insight into this process, we systematically analysed the interactions between Pr55^Gag or its maturation products (NCp15, NCp9 and NCp7) and the 5' gRNA region and their structural consequences, in vitro. We show that Pr55^Gag and its maturation products mostly bind at different RNA sites and with different contributions of their two zinc knuckle domains. Importantly, these proteins have different transient and permanent effects on the RNA structure, the late NCp9 and NCp7 inducing dramatic structural rearrangements. Altogether, our results reveal the distinct contributions of the different Pr55^Gag maturation products on the gRNA structural maturation.

SUBMITTER: Gilmer O

PROVIDER: S-EPMC8786341 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55<sup>Gag</sup> and its maturation products.

Gilmer Orian O Mailler Elodie E Paillart Jean-Christophe JC Mouhand Assia A Tisné Carine C Mak Johnson J Smyth Redmond P RP Marquet Roland R Vivet-Boudou Valérie V

RNA biology 20211231 1

Maturation of the HIV-1 viral particles shortly after budding is required for infectivity. During this process, the Pr55<sup>Gag</sup> precursor undergoes a cascade of proteolytic cleavages, and whilst the structural rearrangements of the viral proteins are well understood, the concomitant maturation of the genomic RNA (gRNA) structure is unexplored, despite evidence that it is required for infectivity. To get insight into this process, we systematically analysed the interactions between Pr55<su ...[more]

PMID: 35067194

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Project description:Late in the HIV-1 replication cycle, the viral structural protein Gag is targeted to virus assembly sites at the plasma membrane of infected cells. The capsid (CA) domain of Gag plays a critical role in the formation of the hexameric Gag lattice in the immature virion, and, during particle release, CA is cleaved from the Gag precursor by the viral protease and forms the conical core of the mature virion. A highly conserved Pro-Pro-Ile-Pro (PPIP) motif (CA residues 122 to 125) [PPIP(122-125)] in a loop connecting CA helices 6 and 7 resides at a 3-fold axis formed by neighboring hexamers in the immature Gag lattice. In this study, we characterized the role of this PPIP(122-125) loop in HIV-1 assembly and maturation. While mutations P123A and P125A were relatively well tolerated, mutation of P122 and I124 significantly impaired virus release, caused Gag processing defects, and abolished infectivity. X-ray crystallography indicated that the P122A and I124A mutations induce subtle changes in the structure of the mature CA lattice which were permissive for in vitro assembly of CA tubes. Transmission electron microscopy and cryo-electron tomography demonstrated that the P122A and I124A mutations induce severe structural defects in the immature Gag lattice and abrogate conical core formation. Propagation of the P122A and I124A mutants in T-cell lines led to the selection of compensatory mutations within CA. Our findings demonstrate that the CA PPIP(122-125) loop comprises a structural element critical for the formation of the immature Gag lattice.IMPORTANCE Capsid (CA) plays multiple roles in the HIV-1 replication cycle. CA-CA domain interactions are responsible for multimerization of the Gag polyprotein at virus assembly sites, and in the mature virion, CA monomers assemble into a conical core that encapsidates the viral RNA genome. Multiple CA regions that contribute to the assembly and release of HIV-1 particles have been mapped and investigated. Here, we identified and characterized a Pro-rich loop in CA that is important for the formation of the immature Gag lattice. Changes in this region disrupt viral production and abrogate the formation of infectious, mature virions. Propagation of the mutants in culture led to the selection of second-site compensatory mutations within CA. These results expand our knowledge of the assembly and maturation steps in the viral replication cycle and may be relevant for development of antiviral drugs targeting CA.

Dataset Information

Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55^Gag and its maturation products.

Publications

Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55<sup>Gag</sup> and its maturation products.

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Dataset Information

Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55Gag and its maturation products.

Publications

Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55<sup>Gag</sup> and its maturation products.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Structural maturation of the HIV-1 RNA 5' untranslated region by Pr55^Gag and its maturation products.