In vitro functional analysis of gRNA sites regulating assembly of hepatitis B virus.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
16 12 2021
16 12 2021
Historique:
received:
12
12
2020
accepted:
16
11
2021
entrez:
17
12
2021
pubmed:
18
12
2021
medline:
11
1
2022
Statut:
epublish
Résumé
The roles of RNA sequence/structure motifs, Packaging Signals (PSs), for regulating assembly of an HBV genome transcript have been investigated in an efficient in vitro assay containing only core protein (Cp) and RNA. Variants of three conserved PSs, within the genome of a strain not used previously, preventing correct presentation of a Cp-recognition loop motif are differentially deleterious for assembly of nucleocapsid-like particles (NCPs). Cryo-electron microscopy reconstruction of the T = 4 NCPs formed with the wild-type gRNA transcript, reveal that the interior of the Cp shell is in contact with lower resolution density, potentially encompassing the arginine-rich protein domains and gRNA. Symmetry relaxation followed by asymmetric reconstruction reveal that such contacts are made at every symmetry axis. We infer from their regulation of assembly that some of these contacts would involve gRNA PSs, and confirmed this by X-ray RNA footprinting. Mutation of the ε stem-loop in the gRNA, where polymerase binds in vivo, produces a poor RNA assembly substrate with Cp alone, largely due to alterations in its conformation. The results show that RNA PSs regulate assembly of HBV genomic transcripts in vitro, and therefore may play similar roles in vivo, in concert with other molecular factors.
Identifiants
pubmed: 34916604
doi: 10.1038/s42003-021-02897-2
pii: 10.1038/s42003-021-02897-2
pmc: PMC8677749
doi:
Substances chimiques
RNA, Guide
0
RNA, Viral
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1407Subventions
Organisme : Wellcome Trust
ID : 110145/Z/15/Z
Pays : United Kingdom
Organisme : NIBIB NIH HHS
ID : P30 EB009998
Pays : United States
Organisme : Wellcome Trust
Pays : United Kingdom
Organisme : MRF
ID : MRF_MRF-044-0002-RG-PATEL
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/N021517/1
Pays : United Kingdom
Informations de copyright
© 2021. The Author(s).
Références
J Struct Biol. 2016 Jan;193(1):1-12
pubmed: 26592709
Nat Commun. 2017 Dec;8(1):5
pubmed: 28232749
Genes Dev. 2015 Nov 15;29(22):2349-61
pubmed: 26543158
Cold Spring Harb Perspect Med. 2015 Nov 09;5(12):
pubmed: 26552701
J Virol. 2014 Sep;88(18):10472-9
pubmed: 24965458
Nat Commun. 2012 Apr 17;3:789
pubmed: 22510694
Antiviral Res. 2015 Sep;121:82-93
pubmed: 26129969
Virology. 2007 Mar 15;359(2):371-81
pubmed: 17056086
J Struct Biol. 2012 Dec;180(3):519-30
pubmed: 23000701
Proc Natl Acad Sci U S A. 2012 Sep 25;109(39):15769-74
pubmed: 23019360
EMBO J. 1990 Oct;9(10):3389-96
pubmed: 2209549
Nat Commun. 2019 Jul 16;10(1):3130
pubmed: 31311931
Proc Natl Acad Sci U S A. 2014 Aug 5;111(31):11329-34
pubmed: 25034253
J Virol. 1993 Jun;67(6):3254-63
pubmed: 7684464
Virology. 1985 Jul 15;144(1):181-93
pubmed: 4060587
Nature. 1997 Mar 6;386(6620):91-4
pubmed: 9052787
Methods Enzymol. 2016;579:125-57
pubmed: 27572726
J Comput Chem. 2004 Oct;25(13):1605-12
pubmed: 15264254
N Engl J Med. 1976 Sep 2;295(10):517-22
pubmed: 950957
Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17174-8
pubmed: 17090672
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
pubmed: 20057044
J Virol. 2010 Jul;84(14):7174-84
pubmed: 20427522
Nature. 1997 Mar 6;386(6620):88-91
pubmed: 9052786
Biophys Rev. 2019 Aug;11(4):603-609
pubmed: 31342264
J Viral Hepat. 2000 Jan;7(1):79-83
pubmed: 10718947
J Med Virol. 1987 Oct;23(2):101-7
pubmed: 2445909
Nat Microbiol. 2017 Jun 19;2:17098
pubmed: 28628133
J Viral Hepat. 2006 Apr;13(4):256-63
pubmed: 16611192
Mol Cell. 1999 Jun;3(6):771-80
pubmed: 10394365
Gastroenterology. 2003 Dec;125(6):1714-22
pubmed: 14724824
EMBO J. 1992 Sep;11(9):3413-20
pubmed: 1380455
Microbiol Mol Biol Rev. 2000 Mar;64(1):51-68
pubmed: 10704474
J Synchrotron Radiat. 2021 Sep 1;28(Pt 5):1321-1332
pubmed: 34475281
J Mol Biol. 1997 Feb 14;266(1):144-59
pubmed: 9054977
Virology. 1999 Jul 5;259(2):342-8
pubmed: 10388659
Proc Natl Acad Sci U S A. 2012 Feb 7;109(6):1907-12
pubmed: 22308402
J Synchrotron Radiat. 2019 Jul 1;26(Pt 4):1388-1399
pubmed: 31274468
World J Gastroenterol. 2007 Jan 7;13(1):125-40
pubmed: 17206760
J Virol. 2017 Jul 27;91(16):
pubmed: 28566379
J Mol Biol. 2001 Mar 16;307(1):183-96
pubmed: 11243813
J Mol Biol. 2013 Mar 25;425(6):1050-64
pubmed: 23318955
Virus Res. 2008 Jun;134(1-2):235-49
pubmed: 18339439
Nucleic Acids Res. 2003 Jul 1;31(13):3406-15
pubmed: 12824337
J Virol. 2018 Sep 26;92(20):
pubmed: 30089690
J Mol Recognit. 2006 Nov-Dec;19(6):542-8
pubmed: 17006877
Nature. 2017 Jan 5;541(7635):112-116
pubmed: 27992877
PLoS One. 2013 Sep 06;8(9):e69729
pubmed: 24039702
Viruses. 2020 Dec 23;13(1):
pubmed: 33374798
Nature. 2019 Jan;565(7739):377-381
pubmed: 30626974
J Biol Phys. 2013 Mar;39(2):277-87
pubmed: 23704797
J Mol Biol. 2016 Jan 29;428(2 Pt B):431-48
pubmed: 26608810
Virology. 2010 Nov 25;407(2):281-8
pubmed: 20850162
J Virol. 2000 May;74(10):4721-8
pubmed: 10775610
Proc Natl Acad Sci U S A. 2014 Apr 8;111(14):5361-6
pubmed: 24706827
J Mol Biol. 2011 Oct 14;413(1):51-65
pubmed: 21839093
Proc Natl Acad Sci U S A. 2005 Jun 7;102(23):8138-43
pubmed: 15928089
Proc Natl Acad Sci U S A. 2015 Feb 17;112(7):2227-32
pubmed: 25646435
Antiviral Res. 2016 Jan;125:25-33
pubmed: 26611395
Eur Phys J E Soft Matter. 2006 Mar;19(3):303-10
pubmed: 16554977
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
J Mol Biol. 2013 Sep 9;425(17):3235-49
pubmed: 23763992
Annu Rev Biophys. 2019 May 6;48:495-514
pubmed: 30951648
ACS Infect Dis. 2019 May 10;5(5):759-768
pubmed: 30525438
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
pubmed: 20124702
Elife. 2018 Jan 29;7:
pubmed: 29377794
Elife. 2018 Nov 09;7:
pubmed: 30412051
Nat Rev Microbiol. 2006 Aug;4(8):588-96
pubmed: 16845429
RNA Biol. 2013 Apr;10(4):481-9
pubmed: 23422316
Phys Rev Lett. 2005 Jan 28;94(3):038101
pubmed: 15698326
Virology. 2002 Nov 10;303(1):199-210
pubmed: 12482672
Science. 2021 Jun 11;372(6547):1220-1224
pubmed: 34112695
Epigenetics. 2011 Jun;6(6):720-6
pubmed: 21546797
Nat Methods. 2017 Apr;14(4):331-332
pubmed: 28250466
Phys Rev E Stat Nonlin Soft Matter Phys. 2005 Jun;71(6 Pt 1):061928
pubmed: 16089786
J Virol. 2016 Mar 28;90(8):3994-4004
pubmed: 26842475
PLoS Biol. 2018 Jun 20;16(6):e2006191
pubmed: 29924793
PLoS One. 2015 Mar 17;10(3):e0119820
pubmed: 25782010