Evidence for DNA hairpin recognition by Zta at the Epstein-Barr virus origin of lytic replication.
The Epstein-Barr virus immediate-early protein (Zta) plays an essential role in viral lytic activation and pathogenesis. Zta is a basic zipper (b-Zip) domain-containing protein that binds multiple sites in the viral origin of lytic replication (OriLyt) and is required for lytic-cycle DNA replication. We present evidence that Zta binds to a sequence-specific, imperfect DNA hairpin formed by an inverted repeat within the upstream essential element (UEE) of OriLyt. Mutations in the OriLyt sequence that are predicted to disrupt hairpin formation also disrupt Zta binding in vitro. Restoration of the hairpin rescues the defect. We also show that OriLyt DNA isolated from replicating cells contains a nuclease-sensitive region that overlaps with the inverted-repeat region of the UEE. Furthermore, point mutations in Zta that disrupt specific recognition of the UEE hairpin are defective for activation of lytic replication. These data suggest that Zta acts by inducing and/or stabilizing a DNA hairpin structure during productive infection. The DNA hairpin at OriLyt with which Zta interacts resembles DNA structures formed at other herpesvirus origins and may therefore represent a common secondary structure used by all herpesvirus family members during the initiation of DNA replication.
Rennekamp
Andrew J
AJ
The Wistar Institute, 3601 Spruce St., Philadelphia, PA 19104, USA.
Wang
Pu
P
Lieberman
Paul M
PM
D000595:Amino Acid Sequence; D001483:Base Sequence; D004279:DNA, Viral / Q000378:metabolism*; D020031:Epstein-Barr Virus Infections / Q000235:genetics / Q000378:metabolism; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009154:Mutation; D009690:Nucleic Acid Conformation; D018741:Replication Origin / Q000235:genetics*; D016415:Sequence Alignment; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
C063676:BZLF1 protein, Herpesvirus 4, Human; D004279:DNA, Viral; D015534:Trans-Activators
J Virol. 1997 Apr;71(4):3054-61
9060666
J Virol. 1991 Jun;65(6):3284-92
1851878
J Virol. 2003 Feb;77(4):2477-88
12551986
J Virol. 2005 Nov;79(22):13984-92
16254334
Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7194-9
11416203
J Virol. 1996 Mar;70(3):1759-67
8627698
J Virol. 1996 Nov;70(11):7398-413
8892858
Virology. 1988 Aug;165(2):549-64
2841800
J Virol. 2005 Nov;79(21):13298-309
16227252
J Virol. 2000 Jun;74(12):5726-8
10823882
J Virol. 2009 Aug;83(16):8090-8
19494003
J Virol. 2002 Aug;76(15):7890-6
12097603
J Virol. 1996 Aug;70(8):5131-42
8764021
EMBO J. 1993 Oct;12(10):3921-9
8404860
Annu Rev Biochem. 1997;66:347-84
9242911
Proc Natl Acad Sci U S A. 1994 May 24;91(11):4629-33
8197110
J Virol. 2001 Sep;75(18):8792-802
11507224
Adv Cancer Res. 2007;97:81-109
17419942
Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):2838-42
9096307
Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3896-900
10760262
Mol Cell Biol. 1985 May;5(5):930-42
2987682
EMBO J. 1986 Dec 1;5(12):3243-9
3028777
J Virol. 1996 Feb;70(2):1132-6
8551573
J Virol. 2004 Aug;78(16):8615-29
15280471
J Virol. 1991 May;65(5):2601-11
1850034
J Virol. 1989 Jul;63(7):3109-16
2542618
Proc Natl Acad Sci U S A. 1986 Sep;83(17):6322-6
3018724
J Virol. 1990 Mar;64(3):1143-55
2154599
Nucleic Acids Res. 2008 Jul 1;36(Web Server issue):W70-4
18424795
Virology. 1996 Jun 15;220(2):367-76
8661388
Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10665-9
7938010
J Biol Chem. 2004 Jul 9;279(28):29211-7
15133043
J Virol. 1990 Dec;64(12):6184-95
2173786
J Gen Virol. 1990 Jun;71 ( Pt 6):1379-85
2161905
Proc Natl Acad Sci U S A. 1986 Dec;83(23):9094-8
3024166
J Virol. 1998 Nov;72(11):8559-67
9765394
PLoS Pathog. 2008 Mar 07;4(3):e1000005
18369464
J Virol. 1990 Oct;64(10):4957-67
2168979
Cell Growth Differ. 1992 Aug;3(8):557-63
1327084
J Virol. 2005 Nov;79(22):13993-4003
16254335
J Virol. 1988 Jan;62(1):91-9
2824860
J Gen Virol. 1993 Jul;74 ( Pt 7):1349-55
8393075
J Virol. 2004 Nov;78(21):11664-77
15479808
Proc Natl Acad Sci U S A. 1985 Jun;82(12):4085-9
2987963
J Virol. 1995 May;69(5):2998-3006
7707526
J Virol. 1990 Jul;64(7):3358-69
2161945
J Virol. 1988 Nov;62(11):4096-103
2845124
J Virol. 1992 Nov;66(11):6735-46
1328687
J Biol Chem. 2002 Oct 25;277(43):41204-12
12183471
J Gen Virol. 1986 Aug;67 ( Pt 8):1613-23
3016156
J Virol. 1996 Dec;70(12):8340-7
8970953
Nat Genet. 2004 Oct;36(10):1099-104
15361873
Nat Rev Cancer. 2004 Oct;4(10):757-68
15510157
J Biol Chem. 1999 Oct 1;274(40):28059-62
10497152
Oncogene. 1991 Feb;6(2):195-204
1847997
Proc Natl Acad Sci U S A. 1988 Aug;85(15):5414-8
2840659
Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):898-903
12552114
EMBO J. 1989 Jan;8(1):127-32
2540954
IUBMB Life. 2003 Jan;55(1):13-22
12716057
J Mol Biol. 1996 May 24;258(5):789-99
8637010
Future Virol. 2010 Jan;5(1):65-83
22468146
J Biol Chem. 1990 Oct 5;265(28):17167-73
2170367
J Virol. 2008 May;82(9):4647-55
18305033
J Virol. 1990 Jun;64(6):2560-8
2159531
Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1460-5
9465037
J Virol. 2005 Jan;79(1):245-56
15596820
Herpes. 2000 Oct;7(3):76-82
11867007
PLoS Pathog. 2009 Mar;5(3):e1000356
19325883
Cell. 1987 Nov 6;51(3):473-81
3311385
J Virol. 2005 Jun;79(12):7338-48
15919888
J Virol. 1999 Jun;73(6):4543-51
10233912
J Gen Virol. 2003 Aug;84(Pt 8):1941-1949
12867624
EMBO J. 1996 Apr 1;15(7):1742-50
8612599
J Biol Chem. 1997 Feb 14;272(7):4623-30
9020191
Proc Natl Acad Sci U S A. 1990 Dec;87(23):9459-63
2174563
J Virol. 2006 Nov;80(22):10942-9
16971443
J Virol. 1993 Jul;67(7):4237-45
8389925
Virology. 1992 Oct;190(2):702-15
1325702
J Biol Chem. 2000 Feb 25;275(8):5880-7
10681580
EMBO J. 1982;1(7):863-7
6329712
J Mol Biol. 1975 Nov 5;98(3):503-17
1195397
J Virol. 1992 Aug;66(8):5030-9
1321285