دورية أكاديمية
أعرض في EDS

The interaction of dengue virus capsid protein with negatively charged interfaces drives the in vitro assembly of nucleocapsid-like particles.

التفاصيل البيبلوغرافية
العنوان: The interaction of dengue virus capsid protein with negatively charged interfaces drives the in vitro assembly of nucleocapsid-like particles.
المؤلفون: Mebus-Antunes NC; Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil., Ferreira WS; Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil., Barbosa GM; Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil., Neves-Martins TC; Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil., Weissmuller G; Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil., Almeida FCL; Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil.; Centro Nacional de Biologia Estrutural e Bioimagem, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil., Da Poian AT; Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil.
المصدر: PloS one [PLoS One] 2022 Mar 01; Vol. 17 (3), pp. e0264643. Date of Electronic Publication: 2022 Mar 01 (Print Publication: 2022).
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE
أسماء مطبوعة: Original Publication: San Francisco, CA : Public Library of Science
مواضيع طبية MeSH: Dengue Virus*/genetics, Capsid Proteins/genetics ; Humans ; Nucleocapsid/metabolism ; Oligonucleotides/metabolism ; RNA/metabolism ; Virus Assembly
مستخلص: Dengue virus (DENV) causes a major arthropod-borne viral disease, with 2.5 billion people living in risk areas. DENV consists in a 50 nm-diameter enveloped particle in which the surface proteins are arranged with icosahedral symmetry, while information about nucleocapsid (NC) structural organization is lacking. DENV NC is composed of the viral genome, a positive-sense single-stranded RNA, packaged by the capsid (C) protein. Here, we established the conditions for a reproducible in vitro assembly of DENV nucleocapsid-like particles (NCLPs) using recombinant DENVC. We analyzed NCLP formation in the absence or presence of oligonucleotides in solution using small angle X-ray scattering, Rayleigh light scattering as well as fluorescence anisotropy, and characterized particle structural properties using atomic force and transmission electron microscopy imaging. The experiments in solution comparing 2-, 5- and 25-mer oligonucleotides established that 2-mer is too small and 5-mer is sufficient for the formation of NCLPs. The assembly process was concentration-dependent and showed a saturation profile, with a stoichiometry of 1:1 (DENVC:oligonucleotide) molar ratio, suggesting an equilibrium involving DENVC dimer and an organized structure compatible with NCLPs. Imaging methods proved that the decrease in concentration to sub-nanomolar concentrations of DENVC allows the formation of regular spherical NCLPs after protein deposition on mica or carbon surfaces, in the presence as well as in the absence of oligonucleotides, in this latter case being surface driven. Altogether, the results suggest that in vitro assembly of DENV NCLPs depends on DENVC charge neutralization, which must be a very coordinated process to avoid unspecific aggregation. Our hypothesis is that a specific highly positive spot in DENVC α4-α4' is the main DENVC-RNA binding site, which is required to be firstly neutralized to allow NC formation.
Competing Interests: The authors have declared that no competing interests exist.
References: FEMS Microbiol Rev. 2015 Mar;39(2):155-70. (PMID: 25725010)
Arch Virol. 2009;154(4):695-8. (PMID: 19305942)
J Virol. 2001 Mar;75(5):2119-29. (PMID: 11160716)
J Virol. 2000 May;74(9):4302-9. (PMID: 10756045)
PLoS Pathog. 2020 May 28;16(5):e1008542. (PMID: 32463839)
RNA Biol. 2021 May;18(5):718-731. (PMID: 33406991)
Cell. 1995 Feb 24;80(4):621-30. (PMID: 7867069)
Nat Rev Phys. 2021;3(2):76-91. (PMID: 33728406)
J Appl Crystallogr. 2012 Mar 15;45(Pt 2):342-350. (PMID: 25484842)
Q Rev Biophys. 2003 May;36(2):147-227. (PMID: 14686102)
Virology. 1984 Jan 30;132(2):401-12. (PMID: 6702103)
Science. 2003 Oct 10;302(5643):248. (PMID: 14551429)
Chemistry. 2018 May 23;24(29):7456-7463. (PMID: 29518273)
Cell. 2002 Mar 8;108(5):717-25. (PMID: 11893341)
Sci Rep. 2015 Jul 10;5:10592. (PMID: 26161501)
Nanoscale. 2015 Sep 28;7(36):14953-64. (PMID: 26302823)
FEBS Lett. 2020 Jun;594(12):1989-2004. (PMID: 32510601)
Arch Biochem Biophys. 2019 Mar 30;664:134-148. (PMID: 30742801)
PeerJ. 2016 Nov 9;4:e2670. (PMID: 27867765)
Nat Struct Mol Biol. 2017 Feb;24(2):184-186. (PMID: 28067914)
Biochem Biophys Res Commun. 2005 Sep 2;334(3):901-6. (PMID: 16053919)
Cell Host Microbe. 2019 Nov 13;26(5):606-622.e8. (PMID: 31631053)
Proc Natl Acad Sci U S A. 2018 Nov 6;115(45):11608-11612. (PMID: 30348794)
Microbiol Mol Biol Rev. 2011 Jun;75(2):268-85. (PMID: 21646429)
Methods Mol Biol. 2011;736:31-43. (PMID: 21660719)
Angew Chem Int Ed Engl. 2005 Mar 18;44(13):1965-8. (PMID: 15726554)
J Virol. 2004 Aug;78(15):8078-84. (PMID: 15254179)
Nat Struct Biol. 2003 Nov;10(11):907-12. (PMID: 14528291)
Cell Host Microbe. 2009 Apr 23;5(4):365-75. (PMID: 19380115)
Pathogens. 2020 Jan 05;9(1):. (PMID: 31948047)
Semin Cell Dev Biol. 2018 Jan;73:199-208. (PMID: 28851598)
J Virol. 2014 Jul;88(14):7998-8015. (PMID: 24807709)
Science. 2016 Apr 22;352(6284):467-70. (PMID: 27033547)
Biophys J. 2017 Feb 28;112(4):663-673. (PMID: 28256226)
J Virol. 2002 Nov;76(21):11128-32. (PMID: 12368355)
Nanoscale. 2017 Aug 10;9(31):11327-11337. (PMID: 28762410)
Annu Rev Virol. 2016 Sep 29;3(1):263-281. (PMID: 27501261)
Viruses. 2018 Mar 20;10(3):. (PMID: 29558394)
Biochem J. 2012 Jun 15;444(3):405-15. (PMID: 22428600)
J Virol. 2014 Jan;88(1):477-82. (PMID: 24155405)
Curr Opin Virol. 2021 Apr;47:106-112. (PMID: 33721656)
Nat Struct Mol Biol. 2013 Jan;20(1):105-10. (PMID: 23241927)
J Virol. 1999 Jul;73(7):5309-19. (PMID: 10364277)
Proc Natl Acad Sci U S A. 2004 Mar 9;101(10):3414-9. (PMID: 14993605)
J Virol. 2003 Jun;77(12):7143-9. (PMID: 12768036)
J Virol. 2013 Jul;87(13):7700-7. (PMID: 23637416)
PLoS Pathog. 2009 Oct;5(10):e1000632. (PMID: 19851456)
Anal Chem. 2019 Jan 2;91(1):622-636. (PMID: 30383361)
المشرفين على المادة: 0 (Capsid Proteins)
0 (Oligonucleotides)
63231-63-0 (RNA)
تواريخ الأحداث: Date Created: 20220301 Date Completed: 20220426 Latest Revision: 20231103
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC8887749
DOI: 10.1371/journal.pone.0264643
PMID: 35231063
قاعدة البيانات: MEDLINE
الوصف
تدمد:1932-6203
DOI:10.1371/journal.pone.0264643