CpG methylation controls reactivation of HIV from latency.
DNA methylation of retroviral promoters and enhancers localized in the provirus 5' long terminal repeat (LTR) is considered to be a mechanism of transcriptional suppression that allows retroviruses to evade host immune responses and antiretroviral drugs. However, the role of DNA methylation in the control of HIV-1 latency has never been unambiguously demonstrated, in contrast to the apparent importance of transcriptional interference and chromatin structure, and has never been studied in HIV-1-infected patients. Here, we show in an in vitro model of reactivable latency and in a latent reservoir of HIV-1-infected patients that CpG methylation of the HIV-1 5' LTR is an additional epigenetic restriction mechanism, which controls resistance of latent HIV-1 to reactivation signals and thus determines the stability of the HIV-1 latency. CpG methylation acts as a late event during establishment of HIV-1 latency and is not required for the initial provirus silencing. Indeed, the latent reservoir of some aviremic patients contained high proportions of the non-methylated 5' LTR. The latency controlled solely by transcriptional interference and by chromatin-dependent mechanisms in the absence of significant promoter DNA methylation tends to be leaky and easily reactivable. In the latent reservoir of HIV-1-infected individuals without detectable plasma viremia, we found HIV-1 promoters and enhancers to be hypermethylated and resistant to reactivation, as opposed to the hypomethylated 5' LTR in viremic patients. However, even dense methylation of the HIV-1 5'LTR did not confer complete resistance to reactivation of latent HIV-1 with some histone deacetylase inhibitors, protein kinase C agonists, TNF-alpha, and their combinations with 5-aza-2deoxycytidine: the densely methylated HIV-1 promoter was most efficiently reactivated in virtual absence of T cell activation by suberoylanilide hydroxamic acid. Tight but incomplete control of HIV-1 latency by CpG methylation might have important implications for strategies aimed at eradicating HIV-1 infection.
Blazkova
Jana
J
Institut National de la Santé et de la Recherche Médicale (INSERM), UMR891, Centre de Recherche en Cancérologie de Marseille; and Institut Paoli-Calmettes, Marseille, France.
Trejbalova
Katerina
K
Gondois-Rey
Françoise
F
Halfon
Philippe
P
Philibert
Patrick
P
Guiguen
Allan
A
Verdin
Eric
E
Olive
Daniel
D
Van Lint
Carine
C
Hejnar
Jiri
J
Hirsch
Ivan
I
D000328:Adult; D000368:Aged; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002843:Chromatin / Q000502:physiology; D002999:Clone Cells; D003001:Cloning, Molecular; D018899:CpG Islands; D019175:DNA Methylation; D005260:Female; D015658:HIV Infections / Q000378:metabolism; D016325:HIV Long Terminal Repeat; D018023:HIV Seronegativity; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D008297:Male; D008875:Middle Aged; D011401:Promoter Regions, Genetic; D011533:Proviruses / Q000235:genetics / Q000378:metabolism; D014766:Viremia / Q000235:genetics / Q000378:metabolism; D017735:Virus Latency / Q000502:physiology*
D002843:Chromatin
J Virol. 2006 Jul;80(13):6441-57
16775332
Methods Mol Biol. 2002;200:143-54
11951649
Annu Rev Med. 2009;60:471-84
18947296
J Immunol. 1989 Jan 15;142(2):431-8
2463307
J Clin Immunol. 2001 May;21(3):218-26
11403229
EMBO J. 1990 Apr;9(4):1157-64
2323336
Proc Natl Acad Sci U S A. 2003 Apr 15;100(8):4819-24
12684537
Mol Cell Biol. 2002 May;22(9):2965-73
11940654
EMBO J. 2007 Jan 24;26(2):412-23
17245431
J Virol. 2009 Apr;83(7):3078-93
19144703
Science. 1997 Nov 14;278(5341):1291-5
9360926
Proc Natl Acad Sci U S A. 1998 Aug 18;95(17):10182-7
9707621
J Virol. 2003 Aug;77(15):8227-36
12857891
Blood. 2001 Nov 15;98(10):3006-15
11698284
Cell. 1998 Nov 25;95(5):595-604
9845362
J Virol. 1997 Mar;71(3):2233-40
9032358
Proc Natl Acad Sci U S A. 2009 Apr 14;106(15):6321-6
19336585
J Virol. 2002 Sep;76(17):8776-86
12163598
J Virol. 2005 Jan;79(2):876-83
15613316
J Virol. 2004 Jun;78(12):6122-33
15163705
J Biol Chem. 2006 Jul 21;281(29):19960-8
16687403
EMBO J. 2003 Apr 15;22(8):1868-77
12682019
Exp Cell Res. 2006 Apr 15;312(7):1011-20
16427621
Science. 1999 Jan 29;283(5402):682-6
9924027
Proc Natl Acad Sci U S A. 2001 Jan 16;98(2):565-9
11209056
Nat Med. 2000 Jan;6(1):82-5
10613829
J Virol. 2004 Sep;78(17):9105-14
15308706
Retrovirology. 2005 Oct 22;2:64
16242045
Nature. 2005 Apr 28;434(7037):1093-7
15793563
Retrovirology. 2006 Oct 12;3:69
17034647
Nature. 1987 Apr 16-22;326(6114):711-3
3031512
Science. 1997 Nov 14;278(5341):1295-300
9360927
Blood. 2009 Jan 1;113(1):58-65
18849485
J Clin Invest. 2005 Jan;115(1):128-37
15630452
Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15109-14
10611346
Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16725-30
18936487
J Virol. 2006 Jan;80(2):854-65
16378987
J Infect Dis. 2001 Jan 1;183(1):36-50
11106537
J Virol. 2001 Dec;75(23):11336-43
11689614
Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1908-13
12552096
Nat Immunol. 2006 Mar;7(3):235-9
16482171
J Virol. 2006 Jul;80(14):6801-9
16809286
EMBO J. 2006 Jan 11;25(1):139-49
16319923
Immunity. 2003 Sep;19(3):413-23
14499116
J Virol. 2002 Sep;76(18):9389-97
12186921
Proc Natl Acad Sci U S A. 1981 Dec;78(12):7609-13
6950402
EMBO J. 2004 Jan 14;23(1):138-49
14685282
Nat Med. 1999 May;5(5):512-7
10229227
J Immunol. 1986 Jun 1;136(11):4049-53
2422271
Drug Discov Today. 2005 Feb 1;10(3):197-204
15708534
Nat Med. 2000 Jan;6(1):76-81
10613828
EMBO J. 1999 Nov 1;18(21):6106-18
10545121
EMBO J. 1996 Mar 1;15(5):1112-20
8605881
Biochem Biophys Res Commun. 1990 Apr 16;168(1):141-7
2327994
Nat Med. 1999 Jun;5(6):651-5
10371503
EMBO J. 2001 Apr 2;20(7):1726-38
11285236
EMBO J. 2007 Dec 12;26(24):4985-95
18007589
EMBO J. 2000 Nov 1;19(21):5884-94
11060039
EMBO J. 2007 Jan 24;26(2):424-35
17245432
Virology. 1992 Jun;188(2):451-8
1585628
J Immunol. 1988 Feb 15;140(4):1117-22
2449497
Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):2927-32
9096323
J Virol. 2005 Jun;79(11):6610-9
15890899
J Virol. 2002 Sep;76(18):9103-11
12186894
J Virol. 2004 Oct;78(19):10507-15
15367617
EMBO J. 2006 Apr 19;25(8):1690-9
16601680
J Virol. 2003 Apr;77(7):4025-32
12634362
Nature. 2005 Apr 28;434(7037):1148-52
15793562
Virology. 2003 Jun 20;311(1):157-68
12832213
Virology. 1999 Mar 1;255(1):171-81
10049832
Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3879-84
18332425