Protein kinase CK2 phosphorylation of EB2 regulates its function in the production of Epstein-Barr virus infectious viral particles.
The Epstein-Barr Virus (EBV) early protein EB2 (also called BMLF1, Mta, or SM) promotes the nuclear export of a subset of early and late viral mRNAs and is essential for the production of infectious virions. We show here that in vitro, protein kinase CK2alpha and -beta subunits bind both individually and, more efficiently, as a complex to the EB2 N terminus and that the CK2beta regulatory subunit also interacts with the EB2 C terminus. Immunoprecipitated EB2 has CK2 activity that phosphorylates several sites within the 80 N-terminal amino acids of EB2, including Ser-55, -56, and -57, which are localized next to the nuclear export signal. EB2S3E, the phosphorylation-mimicking mutant of EB2 at these three serines, but not the phosphorylation ablation mutant EB2S3A, efficiently rescued the production of infectious EBV particles by HEK293(BMLF1-KO) cells harboring an EB2-defective EBV genome. The defect of EB2S3A in transcomplementing 293(BMLF1-KO) cells was not due to impaired nucleocytoplasmic shuttling of the mutated protein but was associated with a decrease in the cytoplasmic accumulation of several late viral mRNAs. Thus, EB2-mediated production of infectious EBV virions is regulated by CK2 phosphorylation at one or more of the serine residues Ser-55, -56, and -57.
Medina-Palazon
Cahora
C
INSERM U758, ENS-Lyon, 46 allée d'Italie, 69364 Lyon Cedex 07, France.
Gruffat
Henri
H
Mure
Fabrice
F
Filhol
Odile
O
Vingtdeux-Didier
Valérie
V
Drobecq
Hervé
H
Cochet
Claude
C
Sergeant
Nicolas
N
Sergeant
Alain
A
Manet
Evelyne
E
D000595:Amino Acid Sequence; D047390:Casein Kinase II / Q000737:chemistry* / Q000378:metabolism; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism / Q000821:virology; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000737:chemistry; D015534:Trans-Activators / Q000378:metabolism; D014162:Transfection; D014764:Viral Proteins / Q000378:metabolism
D010750:Phosphoproteins; C090236:SM protein, Human herpesvirus 4; D015534:Trans-Activators; D014764:Viral Proteins; D012694:Serine; D005982:Glutathione Transferase; D047390:Casein Kinase II
Cell. 1991 Jul 26;66(2):383-94
1830244
J Biol Chem. 2007 May 25;282(21):15645-51
17363367
Virology. 1994 Nov 15;205(1):217-27
7975218
J Cell Sci. 1995 Feb;108 ( Pt 2):779-87
7769018
J Biol Chem. 1997 Jul 4;272(27):17191-5
9202041
J Biol Chem. 1997 Sep 26;272(39):24363-70
9305894
Genes Dev. 1998 Mar 15;12(6):868-79
9512520
J Virol. 2000 Jul;74(13):6068-76
10846090
EMBO J. 2000 Jun 15;19(12):3080-9
10856251
J Virol. 2000 Nov;74(21):10142-52
11024143
J Virol. 2000 Dec;74(23):11322-8
11070032
Mol Cell. 2001 Apr;7(4):899-905
11336712
FEBS Lett. 2001 May 4;496(1):44-8
11343704
EMBO J. 2001 Oct 15;20(20):5769-78
11598019
Nature. 2001 Oct 11;413(6856):644-7
11675789
J Biol Chem. 2002 Feb 8;277(6):3935-42
11724776
J Virol. 2002 Oct;76(19):9635-44
12208942
J Cell Sci. 2002 Oct 15;115(Pt 20):3873-8
12244125
J Biol Chem. 2003 Jan 3;278(1):335-42
12403791
Oncogene. 2003 Jan 16;22(2):220-32
12527891
Mol Cell Biol. 2003 Feb;23(3):975-87
12529402
Mol Cell. 2003 Mar;11(3):837-43
12667464
Nucleic Acids Res. 2003 Jul 1;31(13):3635-41
12824383
J Biol Chem. 2003 Sep 26;278(39):37790-8
12857728
Cell. 2004 Apr 30;117(3):311-21
15109492
J Biol Chem. 2004 Jul 30;279(31):33001-11
15155762
J Virol. 2004 Sep;78(17):9412-22
15308735
Nucleic Acids Res. 2004;32(18):5553-69
15486205
Nucleic Acids Res. 1983 Mar 11;11(5):1475-89
6828386
Virology. 1989 Jan;168(1):101-11
2535897
Nucleic Acids Res. 1990 Dec 11;18(23):6835-43
2175879
J Virol. 1998 Dec;72(12):9526-34
9811685
EMBO J. 1999 Feb 15;18(4):1014-24
10022843
J Virol. 1999 Apr;73(4):3246-57
10074178
J Virol. 1999 May;73(5):4090-100
10196305
Biochem Biophys Res Commun. 1999 Apr 21;257(3):731-7
10208852
J Biol Chem. 1999 Oct 8;274(41):28991-8
10506147
J Virol. 2005 Apr;79(7):3949-61
15767397
Biochem J. 2005 Apr 15;387(Pt 2):295-308
15537388
Curr Opin Cell Biol. 2005 Jun;17(3):242-50
15901492
J Virol. 2005 Nov;79(22):14102-11
16254345
J Biol Chem. 2005 Nov 4;280(44):36935-45
16129689
Mol Cell Biol. 2006 Mar;26(5):1631-43
16478985
J Biol Chem. 2006 Oct 20;281(42):31188-201
16895915
J Biol Chem. 2006 Dec 1;281(48):36915-28
17001072
Cell. 2006 Dec 29;127(7):1389-400
17190602
J Virol. 2007 Apr;81(8):4058-69
17287267
Biochemistry. 1991 Nov 19;30(46):11133-40
1932033