Properties and functions of the nucleocapsid protein in virus assembly

Muriaux, Delphine; Darlix, Jean‐Luc

doi:10.4161/rna.7.6.14065

Cited by 110 publications

(124 citation statements)

References 144 publications

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“…Interestingly, this interaction is critical for the HIV-1 LYPX n L-driven budding pathway (26,62,66,67), emphasizing the role of NC in Alix-mediated HIV-1 budding. Several roles have been attributed to NC in the HIV-1 life cycle; they include reverse transcription, integration, trafficking, virus assembly, viral genome encapsidation, and cell-to-cell virus transmission (19,38,44,47,54,58,71). We recently obtained evidence supporting a role for NC in virus budding.…”

mentioning

confidence: 80%

Budding of Retroviruses Utilizing Divergent L Domains Requires Nucleocapsid

Bello

Dussupt

Sette

et al. 2012

J Virol

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eWe recently reported that human immunodeficiency virus type 1 (HIV-1) carrying PTAP and LYPX n L L domains ceased budding when the nucleocapsid (NC) domain was mutated, suggesting a role for NC in HIV-1 release. Here we investigated whether NC involvement in virus release is a property specific to HIV-1 or a general requirement of retroviruses. Specifically, we examined a possible role for NC in the budding of retroviruses relying on divergent L domains and structurally homologous NC domains that harbor diverse protein sequences. We found that NC is critical for the release of viruses utilizing the PTAP motif whether it functions within its native Gag in simian immunodeficiency virus cpzGAB2 (SIVcpzGAB2) or SIVsmmE543 or when it is transplanted into the heterologous Gag protein of equine infectious anemia virus (EIAV). In both cases, virus release was severely diminished even though NC mutant Gag proteins retained the ability to assemble spherical particles. Moreover, budding-defective NC mutants, which displayed particles tethered to the plasma membrane, were triggered to release virus when access to the cell endocytic sorting complex required for transport pathway was restored (i.e., in trans expression of Nedd4.2s). We also examined the role of NC in the budding of EIAV, a retrovirus relying exclusively on the (L)YPX n L-type L domain. We found that EIAV late budding defects were rescued by overexpression of the isolated Alix Bro1 domain (Bro1). Bro1-mediated rescue of EIAV release required the wild-type NC. EIAV NC mutants lost interactions with Bro1 and failed to produce viruses despite retaining the ability to self-assemble. Together, our studies establish a role for NC in the budding of retroviruses harboring divergent L domains and evolutionarily diverse NC sequences, suggesting the utilization of a common conserved mechanism and/or cellular factor rather than a specific motif.

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mentioning

confidence: 80%

Budding of Retroviruses Utilizing Divergent L Domains Requires Nucleocapsid

Bello

Dussupt

Sette

et al. 2012

J Virol

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“…28,29 Deletion of the NC domain of Gag results in the production of viruses defective in Gag-Gag multimerization, virus budding and particle morphogenesis (reviewed in ref. 30). Mutation of the functional domains of NC has shown that the ZFs and basic residues of the HIV and MLV GagNC are involved in gRNA selection and dimerization through tight interactions with the Psi packaging signal.…”

Section: O N O T D I S T R I B U T Ementioning

confidence: 99%

Cell biology of retroviral RNA packaging

et al. 2011

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“…Mature NC results from the protease-mediated cleavage of the Gag polyprotein precursor and plays key roles in HIV-1 replication (2)(3)(4)(5). During the early steps, NC acts as a cofactor of the reverse transcriptase (RT), to promote the initiation of reverse transcription (6)(7)(8), as well as the two obligatory strand transfers (9,10).…”

mentioning

confidence: 99%

The HIV-1 Nucleocapsid Protein Recruits Negatively Charged Lipids To Ensure Its Optimal Binding to Lipid Membranes

Kempf

Postupalenko

Bora

et al. 2015

J Virol

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The HIV-1 Gag polyprotein precursor composed of the matrix (MA), capsid (CA), nucleocapsid (NC), and p6 domains orchestrates virus assembly via interactions between MA and the cell plasma membrane (PM) on one hand and NC and the genomic RNA on the other hand. As the Gag precursor can adopt a bent conformation, a potential interaction of the NC domain with the PM cannot be excluded during Gag assembly at the PM. To investigate the possible interaction of NC with lipid membranes in the absence of any interference from the other domains of Gag, we quantitatively characterized by fluorescence spectroscopy the binding of the mature NC protein to large unilamellar vesicles (LUVs) used as membrane models. We found that NC, either in its free form or bound to an oligonucleotide, was binding with high affinity (ϳ10 7 M ؊1 ) to negatively charged LUVs. The number of NC binding sites, but not the binding constant, was observed to decrease with the percentage of negatively charged lipids in the LUV composition, suggesting that NC and NC/oligonucleotide complexes were able to recruit negatively charged lipids to ensure optimal binding. However, in contrast to MA, NC did not exhibit a preference for phosphatidylinositol-(4,5)-bisphosphate. These results lead us to propose a modified Gag assembly model where the NC domain contributes to the initial binding of the bent form of Gag to the PM. IMPORTANCEThe NC protein is a highly conserved nucleic acid binding protein that plays numerous key roles in HIV-1 replication. While accumulating evidence shows that NC either as a mature protein or as a domain of the Gag precursor also interacts with host proteins, only a few data are available on the possible interaction of NC with lipid membranes. Interestingly, during HIV-1 assembly, the Gag precursor is thought to adopt a bent conformation where the NC domain may interact with the plasma membrane. In this context, we quantitatively characterized the binding of NC, as a free protein or as a complex with nucleic acids, to lipid membranes and showed that the latter constitute a binding platform for NC. Taken together, our data suggest that the NC domain may play a role in the initial binding events of Gag to the plasma membrane during HIV-1 assembly.T he nucleocapsid protein (NC) of human immunodeficiency virus type 1 (HIV-1) is a small basic nucleic acid binding protein with two strictly conserved CCHC motifs that strongly bind zinc (1). Mature NC results from the protease-mediated cleavage of the Gag polyprotein precursor and plays key roles in HIV-1 replication (2-5). During the early steps, NC acts as a cofactor of the reverse transcriptase (RT), to promote the initiation of reverse transcription (6-8), as well as the two obligatory strand transfers (9, 10). Moreover, NC also reduces RT pauses at the initiation step (11-13), increases the overall RT processivity (14, 15), promotes synthesis through pause sites (16-19), helps to remove the RNA fragments resulting from the RT RNase H activity (20), and may contribute to the pro...

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Properties and functions of the nucleocapsid protein in virus assembly

Cited by 110 publications

References 144 publications

Budding of Retroviruses Utilizing Divergent L Domains Requires Nucleocapsid

Budding of Retroviruses Utilizing Divergent L Domains Requires Nucleocapsid

Cell biology of retroviral RNA packaging

The HIV-1 Nucleocapsid Protein Recruits Negatively Charged Lipids To Ensure Its Optimal Binding to Lipid Membranes

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