A Novel Entry/Uncoating Assay Reveals the Presence of at Least Two Species of Viral Capsids During Synchronized HIV-1 Infection.
To better characterize the behavior of HIV-1 capsids we developed EURT, for Entry/Uncoating assay based on core-packaged RNA availability and Translation. EURT is an alternative to Blam-Vpr, but as reporter RNA translation relies on core opening, it can be used to study viral capsids behavior. Our study reveals the existence of two major capsid species, a dead end one in which the viral genome is readily exposed to the cytoplasm and a functional one in which such exposure requires artificial core destabilization. Although reverse transcription drives a faster loss of susceptibility of viral cores to high doses of PF74, it does not lead to higher exposure of the viral genome, implying that viral cores protect the genome irrespectively of reverse transcription. Lastly, IFNα drifts cores from functional to non-functional species, revealing a novel core-destabilizing activity. This assay sheds new light on the behavior of viral cores inside target cells.
CIRI, Centre International de Recherche en Infectiologie, 46 Allée d'Italie, Lyon F69364, France.|INSERM, U1111, 46 Allée d'Italie, Lyon, F69364, France.|Université Claude Bernard Lyon I, 46 Allée d'Italie, Lyon, F69364, France.|CNRS, UMR5308, 46 Allée d'Italie, Lyon, F69364, France.|Ecole Normale Supérieure de Lyon, 46 Allée d'Italie, Lyon, F69364, France.|Université de Lyon, Lyon, France.
Claire
C
Da Silva Santos
CIRI, Centre International de Recherche en Infectiologie, 46 Allée d'Italie, Lyon F69364, France.|INSERM, U1111, 46 Allée d'Italie, Lyon, F69364, France.|Université Claude Bernard Lyon I, 46 Allée d'Italie, Lyon, F69364, France.|CNRS, UMR5308, 46 Allée d'Italie, Lyon, F69364, France.|Ecole Normale Supérieure de Lyon, 46 Allée d'Italie, Lyon, F69364, France.|Université de Lyon, Lyon, France.
Kevin
K
Tartour
CIRI, Centre International de Recherche en Infectiologie, 46 Allée d'Italie, Lyon F69364, France.|INSERM, U1111, 46 Allée d'Italie, Lyon, F69364, France.|Université Claude Bernard Lyon I, 46 Allée d'Italie, Lyon, F69364, France.|CNRS, UMR5308, 46 Allée d'Italie, Lyon, F69364, France.|Ecole Normale Supérieure de Lyon, 46 Allée d'Italie, Lyon, F69364, France.|Université de Lyon, Lyon, France.
Andrea
A
Cimarelli
J Virol. 2006 May;80(10):4683-90
16641261
Cell. 2009 May 1;137(3):433-44
19410541
J Virol. 2010 Sep;84(18):9254-66
20610724
Nature. 2013 Nov 21;503(7476):402-5
24196705
Trends Cell Biol. 2015 Jun;25(6):320-9
25748385
J Virol. 2015 May;89(10):5350-61
25741002
Nature. 2004 Feb 26;427(6977):848-53
14985764
Proc Natl Acad Sci U S A. 2014 Dec 30;111(52):18625-30
25518861
Virology. 2005 Jun 20;337(1):93-101
15882886
Elife. 2014 Dec 17;3:e04114
25517934
J Mol Biol. 1990 Dec 5;216(3):689-99
2124274
J Virol. 2013 Jan;87(1):648-58
23097450
J Virol. 1992 Apr;66(4):2232-9
1548759
J Virol. 2013 Jan;87(1):683-7
23077298
J Virol. 2009 Aug;83(15):7524-35
19457995
J Virol. 2003 Sep;77(17):9295-304
12915545
EMBO J. 2007 Jun 20;26(12):3025-37
17557080
Virology. 2006 Aug 1;351(2):404-19
16650449
Nat Med. 2001 May;7(5):631-4
11329067
J Virol. 2006 Apr;80(8):3712-20
16571788
Structure. 2011 Nov 9;19(11):1573-81
22078557
PLoS Pathog. 2010 Dec 09;6(12):e1001220
21170360
J Virol. 1990 Jul;64(7):3207-11
2191147
Retrovirology. 2015 Oct 29;12:88
26511606
J Virol. 2002 Jun;76(11):5667-77
11991995
Nat Protoc. 2011 Jun;6(6):806-16
21637200
J Virol. 2000 Apr;74(7):3431-9
10755888
J Biol Chem. 2010 Aug 13;285(33):25185-95
20529865
J Virol. 2011 Jan;85(1):542-9
20962083
J Virol. 2016 May 27;90(12 ):5808-23
27076642
J Virol. 1992 Dec;66(12):7176-82
1433510
PLoS Pathog. 2015 Jul 16;11(7):e1005050
26181333
Science. 1999 Jan 1;283(5398):80-3
9872746
Proc Natl Acad Sci U S A. 2006 Apr 4;103(14):5514-9
16540544
Retrovirology. 2014 Dec 11;11:120
25496772
J Virol. 2007 Apr;81(8):4367-70
17267494
Gene Ther. 2006 Jun;13(12):991-4
16525481
J Virol. 2004 Jan;78(2):1026-31
14694135
Proc Natl Acad Sci U S A. 2011 Jun 14;108(24):9975-80
21628558
Front Microbiol. 2015 May 13;6:467
26029198
J Virol. 2000 Feb;74(3):1168-77
10627527
J Virol. 2015 Sep;89(17):9068-79
26109731
Retrovirology. 2013 Jul 09;10:70
23835323
J Virol. 2014 Dec;88(23 ):13613-25
25231297
J Mol Biol. 2012 Mar 30;417(3):224-39
22306406
J Virol. 2010 Jan;84(1):397-406
19846519
J Virol. 2008 Jul;82(13):6557-65
18417568
Virology. 2007 Apr 10;360(2):286-93
17123568
PLoS Pathog. 2016 Jun 20;12(6):e1005709
27322072
Curr Opin Virol. 2014 Feb;4:32-6
24525292
Virology. 2006 Sep 30;353(2):396-409
16828831
Mol Cell Biol. 2000 Jul;20(13):4922-31
10848617
Proc Natl Acad Sci U S A. 2003 Aug 19;100(17):9849-54
12909718
Nat Biotechnol. 2002 Nov;20(11):1151-4
12355096
Methods Mol Biol. 2014;1087:29-36
24158811
EMBO J. 2003 Apr 1;22(7):1707-15
12660176
Nature. 2013 May 30;497(7451):643-6
23719463
Nature. 2011 Jan 20;469(7330):424-7
21248851
Proc Natl Acad Sci U S A. 2014 Feb 18;111(7):2494-9
24550273
Cell Rep. 2015 Nov 24;13(8):1717-31
26586435
J Virol. 2003 Aug;77(15):8541-7
12857923
J Virol. 1994 Nov;68(11):7559-65
7933143
J Virol. 2013 Jan;87(1):422-32
23097435
Retrovirology. 2016 Apr 23;13:28
27107820
PLoS Pathog. 2011 Dec;7(12):e1002439
22174692
J Biol Chem. 2000 Feb 11;275(6):4298-304
10660598
Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10774-9
15249685
J Virol. 2013 Mar;87(5):2587-96
23255800
Cold Spring Harb Perspect Biol. 2012 Oct 01;4(10):null
22815232
PLoS Pathog. 2011 Mar;7(3):e1002009
21455494
PLoS Pathog. 2013;9(6):e1003461
23818857
Nature. 2004 Jul 29;430(6999):569-73
15243629
Viruses. 2014 Nov 04;6(11):4195-211
25375884