Assembly of recombinant human immunodeficiency virus type 1 capsid protein in vitro

Ehrlich, Lorna S.; Agresta, Beth E.; Carter, C A

doi:10.1128/jvi.66.8.4874-4883.1992

Cited by 203 publications

(132 citation statements)

References 52 publications

(54 reference statements)

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“…This manifests itself as a relatively weak complex that is consistent with the observations of monomeric SIV and HIV MAs in solution (Massiah et al, 1994;Matthews et al, 1994Matthews et al, , 1995. The additional driving force necessary for complete virion assembly is provided by oligomerization of the capsid protein (Ehrlich et al, 1992;Dorfman et al, 1994).…”

Section: Implications For Assemblysupporting

confidence: 83%

The solution structure of the bovine leukaemia virus matrix protein and similarity with lentiviral matrix proteins.

et al. 1996

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In the mature virion, retroviral matrix proteins are found in association with the inner face of the viral membrane. They play a critical role in determining the morphogenesis of virus assembly. We have determined the three‐dimensional solution structure of the bovine leukaemia virus (BLV) matrix protein by heteronuclear nuclear magnetic resonance. The protein contains four principal helices that are joined by short, partially structured loops. Despite no sequence similarity with the lentiviruses, the structure shows an intriguing homology with the equivalent protein from the human and simian immunodeficiency viruses. A root‐mean‐square deviation of 3.78 angstrom is observed over the backbone atoms of 36 equivalent helical positions. The similarity implies a possible common assembly unit for the matrix proteins of type C retroviruses.

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Section: Implications For Assemblysupporting

confidence: 83%

The solution structure of the bovine leukaemia virus matrix protein and similarity with lentiviral matrix proteins.

et al. 1996

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“…Accordingly, viral capsids of multiple copies of identical gene products are formed so as to minimize the genetic information required for virus assembly. In many cases, capsid protein dimers serve as building blocks for the assembly of plant and animal viruses (1,6,11,26,39,42,43). Under our experimental conditions, it was possible to resolve at least four discrete N protein oligomers, including dimers, trimers, tetramers, and pentamers.…”

Section: Discussionmentioning

confidence: 95%

Homo-Oligomerization of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein and the Role of Disulfide Linkages

Wootton

Yoo

2003

J Virol

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As a step toward understanding the assembly pathway of the porcine reproductive and respiratory syndrome virus (PRRSV), the oligomeric properties of the nucleocapsid (N) protein were investigated. In this study, we have demonstrated that under nonreducing conditions the N protein forms disulfide-linked homodimers. However, inclusion of an alkylating agent (N-ethylmaleimide [NEM]) prevented disulfide bond formation, suggesting that these intermolecular disulfide linkages were formed as a result of spurious oxidation during cell lysis. In contrast, N protein homodimers isolated from extracellular virions were shown to have formed NEM-resistant intermolecular disulfide linkages, the function of which is probably to impart stability to the virion. Pulse-chase analysis revealed that N protein homodimers become specifically disulfide linked within the virus-infected cell, albeit at the later stages of infection, conceivably when the virus particle buds into the oxidizing environment of the endoplasmic reticulum. Moreover, NEM-resistant disulfide linkages were shown to occur only during productive PRRSV infection, since expression of recombinant N protein did not result in the formation of NEM-resistant disulfide-linked homodimers. Mutational analysis indicated that of the three conserved cysteine residues in the N protein, only the cysteine at position 23 was involved in the formation of disulfide linkages. The N protein dimer was shown to be stable both in the presence and absence of intermolecular disulfide linkages, indicating that noncovalent interactions also play a role in dimerization. Nondisulfide-mediated N protein interactions were subsequently demonstrated both in vitro by the glutathione S-transferase (GST) pull-down assay and in vivo by the mammalian two-hybrid assay. Using a series of N protein deletion mutants fused to GST, amino acids 30 to 37 were shown to be essential for N-N interactions. Furthermore, since RNase A treatment markedly decreased N protein-binding affinity, it appears that at least in vitro, RNA may be involved in bridging N-N interactions. In cross-linking experiments, the N protein was shown to assemble into higher-order structures, including dimers, trimers, tetramers, and pentamers. Together, these findings demonstrate that the N protein possesses self-associative properties, and these likely provide the basis for PRRSV nucleocapsid assembly.Porcine reproductive and respiratory syndrome is a widespread disease causing respiratory disorders in young pigs and reproductive failure in sows and gilts. The disease was first recognized in North America in 1987 (reviewed in reference 15), and shortly thereafter, the etiological agent was isolated from pigs in Europe (45) and in North America (2, 7). North American and European isolates of porcine reproductive and respiratory syndrome virus (PRRSV) correspond to two distinct genotypes (25,27,30), which exhibit significant antigenic differences (31, 46). Consequently, PRRSV has been divided into two subspecies, European (type I) and North Am...

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“…Identification of CA-CA interfaces in the mature HIV-1 capsid CA multimers with the same structural organization as authentic mature capsids have been obtained by selfassembly of CA in vitro, even in the absence of any other biomolecule, and subjected to structural, biochemical and mutational analyses [27][28][29][30][31][32][33][34][35][36][37]. CryoEM studies of mature HIV-1 capsid-like particles assembled in vitro revealed that these are composed of an array of CA hexamers.…”

Section: Identification Of Ca-ca Interfaces In the Immature Hiv-1 Capsidmentioning

confidence: 99%

The capsid protein of human immunodeficiency virus: intersubunit interactions during virus assembly

Mateu

2009

The FEBS Journal

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The capsid protein (CA) of HIV‐1 is composed of two domains, the N‐terminal domain (NTD) and the C‐terminal domain (CTD). During the assembly of the immature HIV‐1 particle, both CA domains constitute a part of the Gag polyprotein, which forms a spherical capsid comprising up to 5000 radially arranged, extended subunits. Gag–Gag interactions in the immature capsid are mediated in large part by interactions between CA domains, which are involved in the formation of a lattice of connected Gag hexamers. After Gag proteolysis during virus maturation, the CA protein is released, and approximately 1000–1500 free CA subunits self‐assemble into a truncated cone‐shaped capsid. In the mature capsid, NTD–NTD and NTD–CTD interfaces are involved in the formation of CA hexamers, and CTD–CTD interfaces connect neighboring hexamers through homodimerization. The CA–CA interfaces involved in the assembly of the immature capsid and those forming the mature capsid are different, at least in part. CA appears to have evolved an extraordinary conformational plasticity, which allows the creation of multiple CA–CA interfaces and the occurrence of CA conformational switches. This minireview focuses on recent structure–function studies of the diverse CA–CA interactions and interfaces involved in HIV‐1 assembly. Those studies are leading to a better understanding of molecular recognition events during virus morphogenesis, and are also relevant for the development of anti‐HIV drugs that are able to interfere with capsid assembly or disassembly.

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Assembly of recombinant human immunodeficiency virus type 1 capsid protein in vitro

Cited by 203 publications

References 52 publications

The solution structure of the bovine leukaemia virus matrix protein and similarity with lentiviral matrix proteins.

The solution structure of the bovine leukaemia virus matrix protein and similarity with lentiviral matrix proteins.

Homo-Oligomerization of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein and the Role of Disulfide Linkages

The capsid protein of human immunodeficiency virus: intersubunit interactions during virus assembly

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