Potential Role for CA-SP in Nucleating Retroviral Capsid Maturation

England, Matthew R.; Purdy, John; Ropson, Ira J.; Dalessio, Paula M.; Craven, Rebecca

doi:10.1128/jvi.00309-14

Cited by 5 publications

(12 citation statements)

References 63 publications

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“…Additionally, removal of the salt-bridge network through alanine mutations resulted in a collapsed bundle in a 200 ns MD simulation (Figure S4C). Importantly, the refined 6HB model is consistent with the findings that removal of charge by mutations R493A and R495A disrupts Gag assembly in vitro (Yu et al, 2001) and that the addition of the first 8 NC residues to purified CA-SP shifts its assembly mode from mature, or CA-like, to a more Gag-like behavior (England et al, 2014). Such a charged motif has been proposed to assist in the formation of helical bundles in the murine leukemia virus (MLV) via directed charged surfaces (Cheslock et al, 2003).…”

Section: Discussionsupporting

confidence: 77%

“…While the SP subdomain is absent in the Mason-Pfizer monkey virus (M-PMV), this subdomain is a necessary component for Gag assembly of both RSV and HIV-1 (Craven et al, 1993; Kräusslich et al, 1995; Wright et al, 2007; Datta et al, 2011) as it provides crucial lateral stabilization of hexameric Gag complexes (von Schwedler et al, 2003; Li et al, 2007; Larson et al, 2003). Mutations or deletions of SP/SP1 residues interfere with Gag-Gag interactions in vitro (Keller et al, 2008; Bush et al, 2014; England et al, 2014; Datta et al, 2011; Yu et al, 2001) and cause in vivo phenotypes ranging from mild to severe disturbances of virion morphology and even to the total blockage of virus assembly (Kräusslich et al, 1995; Liang et al, 2003; Morikawa et al, 2000). …”

Section: Introductionmentioning

confidence: 99%

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Atomic Modeling of an Immature Retroviral Lattice Using Molecular Dynamics and Mutagenesis

et al. 2015

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Defining the molecular interaction between Gag proteins in an assembled hexagonal lattice of immature retrovirus particles is crucial for elucidating the mechanisms of virus assembly and maturation. Recent advances in cryo-electron microscopy have yielded subnanometer structural information on the morphology of immature Gag lattices, making computational modeling and simulations feasible for investigating the Gag-Gag interactions at the atomic level. We have examined the structure of Rous sarcoma virus (RSV) using all-atom molecular dynamics simulations and in vitro assembly, to create the first all-atom model of an immature retroviral lattice. Microseconds-long replica exchange molecular dynamics simulation of the spacer peptide (SP)-nucleocapsid (NC) subdomains results in a 6-helix bundle with amphipathic properties. The resulting model of the RSV Gag lattice shows features and dynamics of the capsid protein (CA) with implications for the maturation process, and confirms the stabilizing role of the upstream and downstream regions of Gag, namely p10 and SP-NC.

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Section: Discussionsupporting

confidence: 77%

Section: Introductionmentioning

confidence: 99%

Atomic Modeling of an Immature Retroviral Lattice Using Molecular Dynamics and Mutagenesis

et al. 2015

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“…To follow tube assembly in a more quantitative fashion we used a turbidimetric assay, in which the attenuation of incident light due to scattering by the tubes was measured using a conventional absorption spectrophotometer. Similar assays have been used to study HIV-1 CA tube formation 56,58,62,[66][67][68][69] as well as the phosphate-driven assembly of RSV CA into CLPs 50,52 . For long, thin tubes (relative to the wavelength of the incident light) the turbidity is a direct measure of the mass of protein that has polymerized 70,71 .…”

Section: Development Of a Turbidimetric Assay For Tube Assembly And Dmentioning

confidence: 99%

In vitroassembly of the Rous Sarcoma Virus capsid protein into hexamer tubes under physiological conditions

Jaballah

Bailey

Desfosses

et al. 2017

Preprint

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During a proteolytically-driven maturation process, the ortho-retroviral capsid protein (CA) assembles to form the convex shell that surrounds the viral genome. In some orthoretroviruses, including Rous Sarcoma Virus (RSV), CA carries a short and hydrophobic spacer peptide (SP) at its C-terminus early in the maturation process, which is progressively removed as maturation proceeds. In this work, we show that RSV CA assembles in vitro at physiological temperatures, forming hexamer tubes that effectively model the mature capsid surface. Tube assembly is strongly influenced by electrostatic effects, and is a nucleated process that remains thermodynamically favored at lower temperatures, but is effectively arrested by the large Gibbs energy barrier associated with nucleation.

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“…The gray rectangle marks the experimentally confirmed helical part of the region. rangement of capsid protein from its immature to mature form (38).…”

Section: Figure 6 Potential Intrahelical Ionic Interactions Stabilizmentioning

confidence: 99%

Functional and Structural Characterization of Novel Type of Linker Connecting Capsid and Nucleocapsid Protein Domains in Murine Leukemia Virus

Doležal

Hadravová

Kožíšek

et al. 2016

Journal of Biological Chemistry

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The assembly of immature retroviral particles is initiated in the cytoplasm by the binding of the structural polyprotein precursor Gag with viral genomic RNA. The protein interactions necessary for assembly are mediated predominantly by the capsid (CA) and nucleocapsid (NC) domains, which have conserved structures. In contrast, the structural arrangement of the CA-NC connecting region differs between retroviral species. In HIV-1 and Rous sarcoma virus, this region forms a rod-like structure that separates the CA and NC domains, whereas in Mason-Pfizer monkey virus, this region is densely packed, thus holding the CA and NC domains in close proximity. Interestingly, the sequence connecting the CA and NC domains in gammaretroviruses, such as murine leukemia virus (MLV), is unique. The sequence is called a charged assembly helix (CAH) due to a high number of positively and negatively charged residues. Although both computational and deletion analyses suggested that the MLV CAH forms a helical conformation, no structural or biochemical data supporting this hypothesis have been published. Using an in vitro assembly assay, alanine scanning mutagenesis, and biophysical techniques (circular dichroism, NMR, microcalorimetry, and electrophoretic mobility shift assay), we have characterized the structure and function of the MLV CAH. We provide experimental evidence that the MLV CAH belongs to a group of charged, E(R/K)-rich, single ␣-helices. This is the first single ␣-helix motif identified in viral proteins.

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Potential Role for CA-SP in Nucleating Retroviral Capsid Maturation

Abstract: During virion maturation, the Rous sarcoma virus (RSV) capsid protein is cleaved from the

Cited by 5 publications

References 63 publications

Atomic Modeling of an Immature Retroviral Lattice Using Molecular Dynamics and Mutagenesis

Atomic Modeling of an Immature Retroviral Lattice Using Molecular Dynamics and Mutagenesis

In vitroassembly of the Rous Sarcoma Virus capsid protein into hexamer tubes under physiological conditions

Functional and Structural Characterization of Novel Type of Linker Connecting Capsid and Nucleocapsid Protein Domains in Murine Leukemia Virus

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