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Effect of Small Polyanions on In Vitro Assembly of Selected Members of Alpha-, Beta- and Gammaretroviruses

. 2021 Jan 18 ; 13 (1) : . [epub] 20210118

Language English Country Switzerland Media electronic

Document type Journal Article, Research Support, Non-U.S. Gov't

The assembly of a hexameric lattice of retroviral immature particles requires the involvement of cell factors such as proteins and small molecules. A small, negatively charged polyanionic molecule, myo-inositol hexaphosphate (IP6), was identified to stimulate the assembly of immature particles of HIV-1 and other lentiviruses. Interestingly, cryo-electron tomography analysis of the immature particles of two lentiviruses, HIV-1 and equine infectious anemia virus (EIAV), revealed that the IP6 binding site is similar. Based on this amino acid conservation of the IP6 interacting site, it is presumed that the assembly of immature particles of all lentiviruses is stimulated by IP6. Although this specific region for IP6 binding may be unique for lentiviruses, it is plausible that other retroviral species also recruit some small polyanion to facilitate the assembly of their immature particles. To study whether the assembly of retroviruses other than lentiviruses can be stimulated by polyanionic molecules, we measured the effect of various polyanions on the assembly of immature virus-like particles of Rous sarcoma virus (RSV), a member of alpharetroviruses, Mason-Pfizer monkey virus (M-PMV) representative of betaretroviruses, and murine leukemia virus (MLV), a member of gammaretroviruses. RSV, M-PMV and MLV immature virus-like particles were assembled in vitro from truncated Gag molecules and the effect of selected polyanions, myo-inostol hexaphosphate, myo-inositol, glucose-1,6-bisphosphate, myo-inositol hexasulphate, and mellitic acid, on the particles assembly was quantified. Our results suggest that the assembly of immature particles of RSV and MLV was indeed stimulated by the presence of myo-inostol hexaphosphate and myo-inositol, respectively. In contrast, no effect on the assembly of M-PMV as a betaretrovirus member was observed.

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Kutluay S.B., Bieniasz P.D. Analysis of the Initiating Events in HIV-1 Particle Assembly and Genome Packaging. PLoS Pathog. 2010;6:e1001200. doi: 10.1371/journal.ppat.1001200. PubMed DOI PMC

Jouvenet N., Simon S.M., Bieniasz P.D. Imaging the interaction of HIV-1 genomes and Gag during assembly of individual viral particles. Proc. Natl. Acad. Sci. USA. 2009;106:19114–19119. doi: 10.1073/pnas.0907364106. PubMed DOI PMC

Schur F.K.M., Hagen W.J.H., Rumlová M., Ruml T., Müller B., Kräusslich H.-G., Briggs J.A.G. Structure of the immature HIV-1 capsid in intact virus particles at 8.8 Å resolution. Nat. Cell Biol. 2015;517:505–508. doi: 10.1038/nature13838. PubMed DOI

Qu K., Glass B., Doležal M., Schur F.K.M., Murciano B., Rein A., Rumlová M., Ruml T., Kräusslich H.-G., Briggs J.A. Structure and architecture of immature and mature murine leukemia virus capsids. Proc. Natl. Acad. Sci. USA. 2018;115:E11751–E11760. doi: 10.1073/pnas.1811580115. PubMed DOI PMC

Bharat T.A.M., Davey N.E., Ulbrich P., Riches J.D., De Marco A., Rumlova M., Sachse C., Ruml T., Briggs J.A.G. Structure of the immature retroviral capsid at 8 Å resolution by cryo-electron microscopy. Nat. Cell Biol. 2012;487:385–389. doi: 10.1038/nature11169. PubMed DOI

Schur F.K.M., Dick R.A., Hagen W.J.H., Vogt V.M., Briggs J.A.G. The Structure of Immature Virus-Like Rous Sarcoma Virus Gag Particles Reveals a Structural Role for the p10 Domain in Assembly. J. Virol. 2015;89:10294–10302. doi: 10.1128/JVI.01502-15. PubMed DOI PMC

Bryant M., Ratner L. Myristoylation-dependent replication and assembly of human immunodeficiency virus. Proc. Natl. Acad. Sci. USA. 1990;87:523–527. doi: 10.1073/pnas.87.2.523. PubMed DOI PMC

Zhou W., Parent L.J., Wills J.W., Resh M.D. Identification of a membrane-binding domain within the amino-terminal region of human immunodeficiency virus type 1 Gag protein which interacts with acidic phospholipids. J. Virol. 1994;68:2556–2569. doi: 10.1128/JVI.68.4.2556-2569.1994. PubMed DOI PMC

Saad J.S., Miller J., Tai J., Kim A., Ghanam R.H., Summers M.F. Structural basis for targeting HIV-1 Gag proteins to the plasma membrane for virus assembly. Proc. Natl. Acad. Sci. USA. 2006;103:11364–11369. doi: 10.1073/pnas.0602818103. PubMed DOI PMC

Ono A., Ablan S.D., Lockett S.J., Nagashima K., Freed E.O. Phosphatidylinositol (4,5) bisphosphate regulates HIV-1 Gag targeting to the plasma membrane. Proc. Natl. Acad. Sci. USA. 2004;101:14889–14894. doi: 10.1073/pnas.0405596101. PubMed DOI PMC

Rhee S.S., Hunter E. Myristylation is required for intracellular transport but not for assembly of D-type retrovirus capsids. J. Virol. 1987;61:1045–1053. doi: 10.1128/JVI.61.4.1045-1053.1987. PubMed DOI PMC

Rhee S.S., Hunter E. Amino acid substitutions within the matrix protein of type D retroviruses affect assembly, transport and membrane association of a capsid. EMBO J. 1991;10:535–546. doi: 10.1002/j.1460-2075.1991.tb07980.x. PubMed DOI PMC

McDonnell J.M., Fushman D., Cahill S.M., Zhou W., Wolven A., Wilson C.B., Nelle T.D., Resh M.D., Wills J., Cowburn D. Solution structure and dynamics of the bioactive retroviral M domain from rous sarcoma virus. J. Mol. Biol. 1998;279:921–928. doi: 10.1006/jmbi.1998.1788. PubMed DOI

Dick R.A., Kamynina E., Vogt V.M. Effect of Multimerization on Membrane Association of Rous Sarcoma Virus and HIV-1 Matrix Domain Proteins. J. Virol. 2013;87:13598–13608. doi: 10.1128/JVI.01659-13. PubMed DOI PMC

Li F., Jin J., Herrmann C., Mothes W. Basic Residues in the Matrix Domain and Multimerization Target Murine Leukemia Virus Gag to the Virological Synapse. J. Virol. 2013;87:7113–7126. doi: 10.1128/JVI.03263-12. PubMed DOI PMC

Bush D.L., Vogt V.M. In Vitro Assembly of Retroviruses. Annu. Rev. Virol. 2014;1:561–580. doi: 10.1146/annurev-virology-031413-085427. PubMed DOI

Krausslich H.G., Facke M., Heuser A.M., Konvalinka J., Zentgraf H. The spacer peptide between human immunodefi-ciency virus capsid and nucleocapsid proteins is essential for ordered assembly and viral infectivity. J. Virol. 1995;69:3407–3419. doi: 10.1128/JVI.69.6.3407-3419.1995. PubMed DOI PMC

Datta S.A., Temeselew L.G., Crist R.M., Soheilian F., Kamata A., Mirro J., Harvin D., Nagashima K., Cachau R.E., Rein A. On the Role of the SP1 Domain in HIV-1 Particle Assembly: A Molecular Switch? J. Virol. 2011;85:4111–4121. doi: 10.1128/JVI.00006-11. PubMed DOI PMC

Wright E.R., Schooler J.B., Ding H.J., Kieffer C., Fillmore C., Sundquist W.I., Jensen G.J. Electron cryotomography of immature HIV-1 virions reveals the structure of the CA and SP1 Gag shells. EMBO J. 2007;26:2218–2226. doi: 10.1038/sj.emboj.7601664. PubMed DOI PMC

Gross I., Hohenberg H., Wilk T., Wiegers K., Grättinger M., Müller B., Fuller S., Kräusslich H. A conformational switch controlling HIV-1 morphogenesis. EMBO J. 2000;19:103–113. doi: 10.1093/emboj/19.1.103. PubMed DOI PMC

Schur F.K.M., Obr M., Hagen W.J.H., Wan W., Jakobi A.J., Kirkpatrick J.M., Sachse C., Kräusslich H.-G., Briggs J.A. An atomic model of HIV-1 capsid-SP1 reveals structures regulating assembly and maturation. Science. 2016;353:506–508. doi: 10.1126/science.aaf9620. PubMed DOI

Keller P.W., Johnson M.C., Vogt V.M. Mutations in the Spacer Peptide and Adjoining Sequences in Rous Sarcoma Virus Gag Lead to Tubular Budding. J. Virol. 2008;82:6788–6797. doi: 10.1128/JVI.00213-08. PubMed DOI PMC

Bush D.L., Monroe E.B., Bedwell G.J., Prevelige P.E., Phillips J.M., Vogt V.M. Higher-Order Structure of the Rous Sarcoma Virus SP Assembly Domain. J. Virol. 2014;88:5617–5629. doi: 10.1128/JVI.02659-13. PubMed DOI PMC

Doležal M., Hadravová R., Kožíšek M., Bednárová L., Langerová H., Ruml T., Rumlová M. Functional and Structural Characterization of Novel Type of Linker Connecting Capsid and Nucleocapsid Protein Domains in Murine Leukemia Virus. J. Biol. Chem. 2016;291:20630–20642. doi: 10.1074/jbc.M116.746461. PubMed DOI PMC

Bohmová K., Hadravová R., Štokrová J., Tůma R., Ruml T., Pichová I., Rumlová M. Effect of Dimerizing Domains and Basic Residues on In Vitro and In Vivo Assembly of Mason-Pfizer Monkey Virus and Human Immunodeficiency Virus. J. Virol. 2009;84:1977–1988. doi: 10.1128/JVI.02022-09. PubMed DOI PMC

Strohalmová-Bohmová K., Spiwok V., Lepšík M., Hadravová R., Křížová I., Ulbrich P., Pichová I., Bednárová L., Ruml T., Rumlová M. Role of Mason-Pfizer Monkey Virus CA-NC Spacer Peptide-Like Domain in Assembly of Immature Particles. J. Virol. 2014;88:14148–14160. doi: 10.1128/JVI.02286-14. PubMed DOI PMC

Cheslock S.R., Poon D.T.K., Fu W., Rhodes T.D., Henderson L.E., Nagashima K., McGrath C.F., Hu W.-S. Charged assembly helix motif in murine leukemia virus capsid: An important region for virus assembly and particle size determination. J. Virol. 2003;77:7058–7066. doi: 10.1128/JVI.77.12.7058-7066.2003. PubMed DOI PMC

De Marco A., Davey N.E., Ulbrich P., Phillips J.M., Lux V., Riches J., Fuzik T., Ruml T., Kräusslich H.-G., Vogt V.M., et al. Conserved and Variable Features of Gag Structure and Arrangement in Immature Retrovirus Particles. J. Virol. 2010;84:11729–11736. doi: 10.1128/JVI.01423-10. PubMed DOI PMC

Füzik T., Píchalová R., Schur F.K.M., Strohalmová K., Křížová I., Hadravová R., Rumlová M., Briggs J.A.G., Ulbrich P., Ruml T. Nucleic Acid Binding by Mason-Pfizer Monkey Virus CA Promotes Virus Assembly and Genome Packaging. J. Virol. 2016;90:4593–4603. doi: 10.1128/JVI.03197-15. PubMed DOI PMC

Mouland A.J., Mercier J., Luo M., Bernier L., DesGroseillers L., Cohen E.A. The Double-Stranded RNA-Binding Protein Staufen Is Incorporated in Human Immunodeficiency Virus Type 1: Evidence for a Role in Genomic RNA Encapsidation. J. Virol. 2000;74:5441–5451. doi: 10.1128/JVI.74.12.5441-5451.2000. PubMed DOI PMC

Joshi A., Garg H., Nagashima K., Bonifacino J.S., Freed E.O. GGA and Arf Proteins Modulate Retrovirus Assembly and Release. Mol. Cell. 2008;30:227–238. doi: 10.1016/j.molcel.2008.03.015. PubMed DOI PMC

Zimmerman C., Klein K.C., Kiser P.K., Singh A.R., Firestein B.L., Riba S.C., Lingappa J.R. Identification of a host protein essential for assembly of immature HIV-1 capsids. Nat. Cell Biol. 2002;415:88–92. doi: 10.1038/415088a. PubMed DOI

Dick R.A., Zadrozny K.K., Xu C., Schur F.K.M., Lyddon T.D., Ricana C.L., Wagner J.M., Perilla J.R., Gan-ser-Pornillos B.K., Johnson M.C., et al. Inositol phosphates are assembly co-factors for HIV. Nature. 2018;560:509–512. doi: 10.1038/s41586-018-0396-4. PubMed DOI PMC

Datta S.A.K., Zhao Z., Clark P.K., Tarasov S., Alexandratos J.N., Campbell S.J., Kvaratskhelia M., Lebowitz J., Rein A. Interactions between HIV-1 Gag Molecules in Solution: An Inositol Phosphate-mediated Switch. J. Mol. Biol. 2007;365:799–811. doi: 10.1016/j.jmb.2006.10.072. PubMed DOI PMC

Dick R.A., Xu C., Morado D.R., Kravchuk V.O., Ricana C., Lyddon T.D., Broad A.M., Feathers J.R., Johnson M.C., Vogt V.M., et al. Structures of immature EIAV Gag lattices reveal a conserved role for IP6 in lentivirus assembly. PLoS Pathog. 2020;16:e1008277. doi: 10.1371/journal.ppat.1008277. PubMed DOI PMC

Hadravová R., Rumlová M., Ruml T. FAITH—Fast Assembly Inhibitor Test for HIV. Virology. 2015;486:78–87. doi: 10.1016/j.virol.2015.08.029. PubMed DOI

Dostálková A., Kaufman F., Křížová I., Vokatá B., Ruml T., Rumlová M. In Vitro Quantification of the Effects of IP6 and Other Small Polyanions on Immature HIV-1 Particle Assembly and Core Stability. J. Virol. 2020;94 doi: 10.1128/JVI.00991-20. PubMed DOI PMC

Joshi S.M., Vogt V.M. Role of the Rous Sarcoma Virus p10 Domain in Shape Determination of Gag Virus-Like Particles Assembled In vitro and within Escherichia coli. J. Virol. 2000;74:10260. doi: 10.1128/JVI.74.21.10260-10268.2000. PubMed DOI PMC

Hadravová R., De Marco A., Ulbrich P., Stokrová J., Dolezal M., Pichová I., Ruml T., Briggs J.A., Rumlová M. In Vitro Assembly of Virus-Like Particles of a Gammaretrovirus, the Murine Leukemia Virus XMRV. J. Virol. 2011;86:1297–1306. doi: 10.1128/JVI.05564-11. PubMed DOI PMC

Klikova M., Rhee S.S., Hunter E., Ruml T. Efficient in vivo and in vitro assembly of retroviral capsids from Gag precursor proteins expressed in bacteria. J. Virol. 1995;69:1093–1098. doi: 10.1128/JVI.69.2.1093-1098.1995. PubMed DOI PMC

Ulbrich P., Haubova S., Nermut M.V., Hunter E., Rumlova M., Ruml T. Distinct roles for nucleic acid in in vitro assem-bly of purified Mason-Pfizer monkey virus CANC proteins. J. Virol. 2006;80:7089–7099. doi: 10.1128/JVI.02694-05. PubMed DOI PMC

Campbell S., Vogt V.M. Self-assembly in vitro of purified CA-NC proteins from Rous sarcoma virus and human immuno-deficiency virus type. J. Virol. 1995;69:6487–6497. doi: 10.1128/JVI.69.10.6487-6497.1995. PubMed DOI PMC

Campbell S., Vogt V.M. In vitro assembly of virus-like particles with Rous sarcoma virus Gag deletion mutants: Identifica-tion of the p10 domain as a morphological determinant in the formation of spherical particles. J. Virol. 1997;71:4425–4435. doi: 10.1128/JVI.71.6.4425-4435.1997. PubMed DOI PMC

Rumlova-Klikova M., Hunter E., Nermut M.V., Pichova I., Ruml T. Analysis of Mason-Pfizer Monkey Virus Gag Domains Required for Capsid Assembly in Bacteria: Role of the N-Terminal Proline Residue of CA in Directing Particle Shape. J. Virol. 2000;74:8452–8459. doi: 10.1128/JVI.74.18.8452-8459.2000. PubMed DOI PMC

Bharat T.A.M., Menendez L.R.C., Hagen W.J.H., Lux V., Igonet S., Schorb M., Schur F.K.M., Kraeusslich H.-G., Briggs J.A. Cryo-electron microscopy of tubular arrays of HIV-1 Gag resolves structures essential for immature virus assembly. Proc. Natl. Acad. Sci. USA. 2014;111:8233–8238. doi: 10.1073/pnas.1401455111. PubMed DOI PMC

Accola M.A., Hoglund S., Gottlinger H.G. A putative alpha-helical structure which overlaps the capsid-p2 boundary in the human immunodeficiency virus type 1 Gag precursor is crucial for viral particle assembly. J. Virol. 1998;72:2072–2078. doi: 10.1128/JVI.72.3.2072-2078.1998. PubMed DOI PMC

Campbell S., Fisher R.J., Towler E.M., Fox S., Issaq H.J., Wolfe T., Phillips L.R., Rein A. Modulation of HIV-like particle assembly in vitro by inositol phosphates. Proc. Natl. Acad. Sci. USA. 2001;98:10875–10879. doi: 10.1073/pnas.191224698. PubMed DOI PMC

Ma Y.M., Vogt V.M. Rous sarcoma virus Gag protein-oligonucleotide interaction suggests a critical role for protein dimer formation in assembly. J. Virol. 2002;76:5452–5462. doi: 10.1128/JVI.76.11.5452-5462.2002. PubMed DOI PMC

Ma Y.M., Vogt V.M. Nucleic acid binding-induced Gag dimerization in the assembly of Rous sarcoma virus particles in vitro. J. Virol. 2004;78:52–60. doi: 10.1128/JVI.78.1.52-60.2004. PubMed DOI PMC

Phillips J.M., Murray P.S., Murray D., Vogt V.M. A molecular switch required for retrovirus assembly participates in the hexagonal immature lattice. EMBO J. 2008;27:1411–1420. doi: 10.1038/emboj.2008.71. PubMed DOI PMC

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