title: The internal Kaposi's sarcoma-associated herpesvirus LANA regions exert a critical role on episome persistence.
abstract: Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a 1,162-amino-acid protein that acts on viral terminal repeat (TR) DNA to mediate KSHV episome persistence. The two essential components of episome persistence are DNA replication prior to cell division and episome segregation to daughter nuclei. These functions are located within N- and C-terminal regions of LANA. N- and C-terminal regions of LANA are sufficient for TR DNA replication. In addition, N- and C-terminal regions of LANA tether episomes to mitotic chromosomes to segregate episomes to progeny cell nuclei. To generate a tethering mechanism, N-terminal LANA binds histones H2A/H2B to attach to mitotic chromosomes, and C-terminal LANA binds TR DNA and also associates with mitotic chromosomes. Here, we test the importance of the internal LANA sequence for episome persistence. We generated LANA mutants that contain N- and C-terminal regions of LANA but have most of the internal sequence deleted. As expected, the LANA mutants bound mitotic chromosomes in a wild-type pattern and also bound TR DNA as assayed by electrophoretic mobility shift assays (EMSA). The mutants mediated TR DNA replication, although with reduced efficiency compared with LANA. Despite the ability to replicate DNA and exert the chromosome and DNA binding functions necessary for segregating episomes to daughter nuclei, the mutants were highly deficient for the ability to mediate both short- and long-term episome persistence. These data indicate that internal LANA sequence exerts a critical effect on its ability to maintain episomes, possibly through effects on TR DNA replication.
authors: Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, 181 Longwood Ave., Boston, MA 02115, USA.
authors: Erika
authors: E
authors: De León Vázquez
authors: Kenneth M
authors: KM
authors: Kaye
references: Science. 1999 Apr 23;284(5414):641-4
references: 10213686
references: J Virol. 2003 Sep;77(18):9758-68
references: 12941884
references: J Virol. 2004 Jul;78(13):7248-56
references: 15194800
references: J Biol Chem. 2003 Feb 28;278(9):7397-405
references: 12486118
references: J Virol. 2004 Jul;78(14):7299-310
references: 15220403
references: PLoS Pathog. 2006 Oct;2(10):e116
references: 17069461
references: Virology. 2001 Dec 20;291(2):241-59
references: 11878894
references: J Virol. 2007 Aug;81(15):8225-35
references: 17522213
references: J Virol. 2003 Jul;77(14):8019-30
references: 12829841
references: J Virol. 2005 Feb;79(4):2637-42
references: 15681465
references: FEBS Lett. 2006 Jan 9;580(1):93-8
references: 16364321
references: J Virol. 2006 Jun;80(11):5273-82
references: 16699007
references: J Biol Chem. 2006 Sep 22;281(38):28113-21
references: 16861237
references: Virology. 2007 Aug 1;364(2):317-29
references: 17434559
references: J Virol. 2007 Feb;81(4):1912-22
references: 17121789
references: J Virol. 2005 Mar;79(6):3468-78
references: 15731241
references: J Virol. 2001 Apr;75(7):3250-8
references: 11238851
references: N Engl J Med. 1995 May 4;332(18):1181-5
references: 7700310
references: Virology. 2004 Feb 20;319(2):225-36
references: 14980483
references: J Gen Virol. 2003 Jun;84(Pt 6):1451-1462
references: 12771414
references: Nature. 1999 Dec 23-30;402(6764):889-94
references: 10622254
references: Blood. 1995 Aug 15;86(4):1276-80
references: 7632932
references: Nat Med. 2000 Oct;6(10):1121-7
references: 11017143
references: J Virol. 2002 Nov;76(22):11596-604
references: 12388720
references: Arch Virol. 2011 Aug;156(8):1323-33
references: 21472413
references: J Biol Chem. 2001 Aug 17;276(33):31016-22
references: 11425857
references: J Virol. 2007 Sep;81(18):9825-37
references: 17626102
references: J Biol Chem. 2002 Jul 26;277(30):27401-11
references: 12015325
references: J Gen Virol. 1999 Nov;80 ( Pt 11):2889-2900
references: 10580050
references: J Virol. 2006 Sep;80(18):8909-19
references: 16940503
references: J Virol. 2004 Oct;78(19):10348-59
references: 15367601
references: J Virol. 2006 Nov;80(21):10772-86
references: 16928766
references: J Mol Biol. 1967 Jun 14;26(2):365-9
references: 4291934
references: Virology. 2006 Jul 20;351(1):18-28
references: 16647097
references: Blood. 2004 Jan 1;103(1):222-8
references: 12969971
references: J Virol. 2005 Nov;79(21):13618-29
references: 16227282
references: J Virol. 2000 Oct;74(20):9637-45
references: 11000236
references: J Gen Virol. 2002 Jan;83(Pt 1):179-188
references: 11752715
references: J Exp Med. 1996 Jul 1;184(1):283-8
references: 8691144
references: Proc Natl Acad Sci U S A. 2006 Sep 26;103(39):14554-9
references: 16983096
references: Virology. 1997 Sep 15;236(1):147-54
references: 9299627
references: J Virol. 2004 Jan;78(1):294-301
references: 14671111
references: Cell Cycle. 2007 Sep 15;6(18):2205-9
references: 17890905
references: J Gen Virol. 2000 Nov;81(Pt 11):2645-2652
references: 11038375
references: Oncogene. 2007 Jul 26;26(34):4979-86
references: 17310999
references: J Virol. 2003 Feb;77(4):2779-83
references: 12552022
references: J Virol. 2009 May;83(9):4326-37
references: 19225000
references: J Virol. 2002 Oct;76(20):10320-31
references: 12239308
references: Virology. 1999 Nov 25;264(2):254-64
references: 10562490
references: J Virol. 2007 Apr;81(8):4348-56
references: 17287261
references: Mol Cancer. 2007 Apr 13;6:28
references: 17433107
references: J Infect Dis. 1999 Nov;180(5):1466-76
references: 10515805
references: Virus Genes. 2007 Apr;34(2):127-36
references: 17143723
references: Nat Med. 2003 Mar;9(3):300-6
references: 12592400
references: J Virol. 1999 Dec;73(12):9789-95
references: 10559289
references: Proc Natl Acad Sci U S A. 2007 Oct 9;104(41):16287-92
references: 17909182
references: J Virol. 2004 Dec;78(24):14033-8
references: 15564510
references: J Virol. 2001 Sep;75(17):7882-92
references: 11483733
references: J Virol. 2004 Sep;78(18):10074-85
references: 15331740
references: Blood. 1995 Oct 1;86(7):2708-14
references: 7670109
references: J Virol. 2006 Jan;80(2):697-709
references: 16378973
references: J Virol. 2009 Nov;83(21):11051-63
references: 19710137
references: J Virol. 1984 Apr;50(1):248-54
references: 6321792
references: J Virol. 2002 Nov;76(22):11677-87
references: 12388727
references: J Gen Virol. 2004 Apr;85(Pt 4):843-855
references: 15039527
references: J Virol. 2007 Oct;81(19):10451-9
references: 17634226
references: J Virol. 2001 Apr;75(8):3948-59
references: 11264383
references: J Virol. 2006 Mar;80(5):2243-56
references: 16474132
references: J Virol. 2010 Oct;84(19):9718-32
references: 20660191
references: J Virol. 2006 Aug;80(16):7965-75
references: 16873253
references: Science. 1994 Dec 16;266(5192):1865-9
references: 7997879
references: Science. 2006 Feb 10;311(5762):856-61
references: 16469929
references: Virology. 2007 Jan 20;357(2):149-57
references: 16979209