Threonine 48 in the BIR domain of survivin is critical to its mitotic and anti-apoptotic activities and can be phosphorylated by CK2 in vitro.
In this study we report that the protein kinase CK2 phosphorylates survivin specifically on threonine 48 (T48) within its BIR domain, and that T48 is critical to both the mitotic and anti-apoptotic roles of survivin. Interestingly, during mitosis T48 mutants localise normally, but are unable to support cell growth when endogenous survivin is removed by siRNA. In addition, while overexpression of survivin normally confers inhibition of TRAIL-mediated apoptosis, this protection is abolished by mutation of T48. Furthermore in interphase cells depletion of endogenous survivin causes redistribution of T48 mutants from the cytoplasm to the nucleus and treatment of cells expressing survivin-GFP with the CK2 inhibitor TBB phenocopies this nuclear redistribution. Finally, we show T48 mutants have increased affinity for borealin, and that this association and cell proliferation can be restored by introduction of a second mutation at T97. To our knowledge these data are the first to identify T48 as a key regulatory site on survivin, and CK2 as a mediator of its mitotic and anti-apoptotic functions.
School of Biomedical Sciences, University of Nottingham, Queen's Medical Centre, Nottingham, UK.
Rachel M A
RM
Barrett
Rita
R
Colnaghi
Sally P
SP
Wheatley
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D017209:Apoptosis; D047390:Casein Kinase II / Q000378:metabolism / Q000502:physiology*; D049109:Cell Proliferation; D049452:Green Fluorescent Proteins / Q000032:analysis; D006367:HeLa Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D008938:Mitosis; D009154:Mutation; D010766:Phosphorylation; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000032:analysis; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein; D000077022:Survivin; D053221:TNF-Related Apoptosis-Inducing Ligand; D013912:Threonine / Q000737:chemistry
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D011993:Recombinant Fusion Proteins; D000077022:Survivin; D053221:TNF-Related Apoptosis-Inducing Ligand; D049452:Green Fluorescent Proteins; D013912:Threonine; D047390:Casein Kinase II
Cell Mol Life Sci. 2002 Dec;59(12):2155-64
12568341
J Cell Biol. 1991 Sep;114(6):1217-32
1894695
Nat Struct Biol. 2000 Jul;7(7):602-8
10876248
Science. 2010 Oct 8;330(6001):231-5
20705812
Curr Biol. 2001 Jun 5;11(11):886-90
11516652
J Biol Chem. 1992 Jul 15;267(20):13943-51
1629192
EMBO J. 1990 Oct;9(10):3253-60
2145148
J Biol Chem. 1991 Oct 25;266(30):20380-9
1939094
J Cell Biol. 1992 Jan;116(1):43-55
1730748
J Biol Chem. 1998 May 1;273(18):11177-82
9556606
Cancer Res. 1999 Dec 15;59(24):6097-102
10626797
J Biol Chem. 2010 Jul 16;285(29):22592-604
20427271
Cell Cycle. 2007 May 15;6(10):1220-30
17457057
J Cell Physiol. 2008 Jun;215(3):750-64
18181150
Proc Natl Acad Sci U S A. 2006 Oct 10;103(41):15079-84
17005722
J Biol Chem. 2006 Jan 13;281(2):1286-95
16291752
Mol Cell. 2001 Sep;8(3):601-11
11583622
J Biol Chem. 1998 Nov 13;273(46):30622-9
9804834
Biochim Biophys Acta. 2008 Jan;1784(1):33-47
17931986
Cancer Res. 2005 May 15;65(10):4362-7
15899828
Oncogene. 2001 Apr 12;20(16):2010-22
11360185
J Biol Chem. 2003 Oct 17;278(42):40514-20
12876286
J Comput Chem. 2004 Oct;25(13):1605-12
15264254
Oncogene. 2003 Jun 5;22(23):3539-47
12789262
Biochem Biophys Res Commun. 2006 Jul 28;346(2):400-7
16762323
FASEB J. 2003 Mar;17(3):349-68
12631575
J Cell Biochem. 2004 Apr 1;91(5):865-79
15034923
Cell Cycle. 2009 Oct 15;8(20):3365-72
19806029
EMBO J. 2003 Jun 16;22(12):2934-47
12805209
Science. 2010 Oct 8;330(6001):235-9
20705815
Mol Cell. 2000 Jul;6(1):183-9
10949039
Nat Rev Mol Cell Biol. 2007 Oct;8(10):798-812
17848966
Cell Cycle. 2007 Mar 1;6(5):518-21
17361097
FEBS Lett. 2001 May 4;496(1):44-8
11343704
Cell Cycle. 2009 Jan 15;8(2):278-83
19158485
Cell Cycle. 2009 Sep 1;8(17):2708-10
19717980
Mol Cell. 2002 Aug;10(2):247-58
12191471
Biochem Biophys Res Commun. 2009 Oct 23;388(3):529-32
19666000
Eur J Biochem. 1990 Apr 30;189(2):251-7
2159876
Mol Cell. 2007 Jul 6;27(1):17-28
17612487
Biochem J. 2001 Sep 15;358(Pt 3):705-15
11535131
Nat Rev Cancer. 2008 Jan;8(1):61-70
18075512
J Biol Chem. 2008 Feb 8;283(6):3289-3296
18057009
Mol Biol Cell. 2004 Mar;15(3):1287-96
14699067
Biochem J. 2002 May 15;364(Pt 1):41-7
11988074
J Biol Chem. 2004 Aug 13;279(33):34087-90
15218035
J Biol Chem. 2007 Nov 30;282(48):35018-23
17881355
Oncogene. 2008 Aug 28;27(37):4986-97
18469858
Trends Cell Biol. 2002 May;12(5):226-30
12062170
Proc Natl Acad Sci U S A. 2000 Nov 21;97(24):13103-7
11069302
Mol Cell Biol. 2009 Apr;29(8):2068-81
19188443
J Cell Sci. 2003 Jul 15;116(Pt 14):2987-98
12783991
Cancer Cell. 2002 Jul;2(1):43-54
12150824
J Biol Chem. 2003 Jul 11;278(28):25277-80
12738781
Cell. 2007 Oct 19;131(2):271-85
17956729
J Biol Chem. 2006 Nov 3;281(44):33450-6
16950794
Cell Cycle. 2008 Apr 15;7(8):1036-46
18414036
J Biol Chem. 2000 Nov 3;275(44):34710-8
10942766
J Biol Chem. 1992 Oct 5;267(28):20317-25
1400350
EMBO J. 2003 Jun 2;22(11):2729-40
12773388
J Biol Chem. 2004 Feb 13;279(7):5655-60
14610074
Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11663-8
16085715
Genome Biol. 2001;2(7):REVIEWS3009
11516343
Cell. 2004 Apr 2;117(1):17-28
15066279
J Cell Biochem. 2001;81(3):445-52
11255227