Human cytomegalovirus IE72 protein interacts with the transcriptional repressor hDaxx to regulate LUNA gene expression during lytic infection.
A putative latency-associated transcript (LUNA) complementary to the human cytomegalovirus (HCMV) UL81-82 region previously identified in seropositive donors' monocytes is also expressed during lytic infection. Thus, the LUNA promoter is active during both lytic and latent infection. Consequently, the mechanisms regulating this promoter may provide further insight into factors that determine whether the outcome of HCMV infection is latent or lytic. By transfection, the LUNA promoter exhibited low but reproducible activity. Substantial activation by virus infection suggested that a viral factor was important for LUNA expression during lytic infection. IE72, a known transactivator of viral promoters, activated the LUNA promoter in cotransfection assays. Furthermore, coinfection with wild-type HCMV but not an IE72 deletion virus (CR208) also activated the LUNA promoter. Finally, diminished LUNA gene expression in CR208 virus-infected cells supported a role for IE72 in LUNA gene expression. The initial regulation of herpesvirus immediate-early gene expression is associated with proteins found at cellular nuclear domain 10 (ND10) bodies, such as PML, hDaxx, and ATRX. hDaxx transfection repressed LUNA promoter activity. Furthermore, we observed binding of hDaxx to the LUNA promoter, which was abrogated by IE72 gene expression via direct interaction. Finally, we show that small interfering RNA (siRNA) knockdown of the hDaxx interaction partner ATRX rescued LUNA gene expression in CR208-infected cells. Overall, these data show that hDaxx/ATRX-mediated repression of LUNA during lytic infection absolutely requires IE72 gene expression. It also suggests that the targeting of cellular factors by IE72 is important throughout the different phases of HCMV gene expression during productive infection.
Reeves
Matthew
M
Department of Medicine, Level 5, Box 157, Addenbrooke's Hospital, Cambridge CB2 2QQ, United Kingdom.
Woodhall
David
D
Compton
Teresa
T
Sinclair
John
J
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D056970:Co-Repressor Proteins; D003587:Cytomegalovirus / Q000235:genetics / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D007231:Infant, Newborn; D018832:Molecular Chaperones; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D017735:Virus Latency / Q000235:genetics
D048868:Adaptor Proteins, Signal Transducing; D056970:Co-Repressor Proteins; C106985:DAXX protein, human; C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D014764:Viral Proteins
J Virol. 2008 Dec;82(24):12543-54
18922870
J Gen Virol. 2005 Nov;86(Pt 11):2949-2954
16227215
Curr Top Microbiol Immunol. 2008;325:117-32
18637503
J Gen Virol. 2007 Nov;88(Pt 11):2935-2940
17947514
Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11321-6
8876134
J Virol. 2002 Aug;76(15):7705-12
12097584
Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):11137-42
8855322
Proc Natl Acad Sci U S A. 2005 Mar 15;102(11):4140-5
15738399
Biochimie. 2007 Jun-Jul;89(6-7):819-30
17343971
J Biol Chem. 2006 Dec 8;281(49):37652-60
17035242
Oncogene. 1999 Jan 28;18(4):935-41
10023669
J Virol. 2008 Nov;82(22):11167-80
18786996
J Virol. 2006 Aug;80(16):8006-18
16873257
J Virol. 1991 Feb;65(2):887-96
1846203
Intervirology. 1996;39(5-6):331-42
9130043
Virology. 2000 Dec 20;278(2):467-76
11118369
Gene. 2006 Dec 15;384:120-8
16989963
Clin Infect Dis. 2004 Jul 15;39(2):233-9
15307033
J Gen Virol. 2009 Oct;90(Pt 10):2364-2374
19515830
J Virol. 2008 Mar;82(6):2661-72
18160441
J Virol. 2004 Feb;78(3):1440-7
14722299
J Virol. 2001 Nov;75(22):10683-95
11602710
J Virol. 2008 Oct;82(19):9525-36
18653451
Exp Cell Res. 1996 Nov 25;229(1):155-8
8940259
EMBO J. 1997 Feb 3;16(3):566-77
9034339
Rev Infect Dis. 1990 Sep-Oct;12 Suppl 7:S701-10
2173099
J Infect Dis. 1988 Aug;158(2):449-56
2841381
J Virol. 2009 Jul;83(13):6391-403
19369332
J Gen Virol. 1998 May;79 ( Pt 5):1233-45
9603339
J Virol. 2006 Jun;80(12):6188-91
16731959
J Virol. 2006 Aug;80(16):8019-29
16873258
J Gen Virol. 1994 Sep;75 ( Pt 9):2337-48
8077932
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13985-90
9391139
J Virol. 2006 Oct;80(20):9998-10009
17005678
Proc Natl Acad Sci U S A. 2003 Sep 16;100(19):10635-40
12953102
Mol Cell Biol. 2000 Mar;20(5):1784-96
10669754
Nucleic Acids Res. 1991 May 11;19(9):2499
2041787
J Gen Virol. 1994 Jun;75 ( Pt 6):1223-33
8207389
J Cell Biol. 1996 Aug;134(4):815-26
8769408
J Virol. 1990 Dec;64(12):6154-65
2173785
J Virol. 2008 Jan;82(1):126-37
17942542
J Virol. 2007 Sep;81(17):9109-20
17596307
J Virol. 2008 Nov;82(21):10922-31
18753203
J Virol. 2003 Jun;77(12):7101-5
12768029
J Virol. 2006 Apr;80(8):3863-71
16571803
J Virol. 2000 Oct;74(19):9333-7
10982383
J Virol. 2004 Jun;78(12):6527-42
15163746
J Virol. 2005 Sep;79(17):11022-34
16103153
J Virol. 1997 Jun;71(6):4599-613
9151854
Rev Infect Dis. 1990 Sep-Oct;12 Suppl 7:S754-66
2173105
J Virol. 2006 Aug;80(16):7995-8005
16873256
J Virol. 2001 Mar;75(5):2388-99
11160742
J Gen Virol. 2006 May;87(Pt 5):1113-1121
16603511
Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7140-5
11390970
J Virol. 1998 Jan;72(1):366-79
9420235
J Virol. 2002 May;76(9):4441-55
11932411
J Cell Sci. 2002 Aug 15;115(Pt 16):3319-30
12140263
Blood. 2007 Aug 1;110(3):937-45
17440050
J Gen Virol. 2006 Jul;87(Pt 7):1763-1779
16760381
Proc Natl Acad Sci U S A. 2004 Dec 7;101(49):17234-9
15572445
J Virol. 2002 Jan;76(2):841-50
11752173
J Virol. 2004 Jul;78(14):7803-12
15220454
Virology. 2000 Aug 15;274(1):39-55
10936087
EMBO J. 2002 Mar 1;21(5):1112-20
11867539