14-3-3ζ regulates the mitochondrial respiratory reserve linked to platelet phosphatidylserine exposure and procoagulant function.
The 14-3-3 family of adaptor proteins regulate diverse cellular functions including cell proliferation, metabolism, adhesion and apoptosis. Platelets express numerous 14-3-3 isoforms, including 14-3-3ζ, which has previously been implicated in regulating GPIbα function. Here we show an important role for 14-3-3ζ in regulating arterial thrombosis. Interestingly, this thrombosis defect is not related to alterations in von Willebrand factor (VWF)-GPIb adhesive function or platelet activation, but instead associated with reduced platelet phosphatidylserine (PS) exposure and procoagulant function. Decreased PS exposure in 14-3-3ζ-deficient platelets is associated with more sustained levels of metabolic ATP and increased mitochondrial respiratory reserve, independent of alterations in cytosolic calcium flux. Reduced platelet PS exposure in 14-3-3ζ-deficient mice does not increase bleeding risk, but results in decreased thrombin generation and protection from pulmonary embolism, leading to prolonged survival. Our studies define an important role for 14-3-3ζ in regulating platelet bioenergetics, leading to decreased platelet PS exposure and procoagulant function.
Heart Research Institute, Thrombosis Group, Newtown, New South Wales 2042, Australia.|Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Simone M
SM
Schoenwaelder
Heart Research Institute, Thrombosis Group, Newtown, New South Wales 2042, Australia.|Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Roxane
R
Darbousset
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Susan L
SL
Cranmer
Centre for Cancer Biology, SA Pathology and the University of South Australia, Adelaide, South Australia 5000, Australia.
Hayley S
HS
Ramshaw
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Stephanie L
SL
Orive
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Sharelle
S
Sturgeon
Heart Research Institute, Thrombosis Group, Newtown, New South Wales 2042, Australia.|Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Yuping
Y
Yuan
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Yu
Y
Yao
Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|School of Life &Environmental Sciences, The University of Sydney, Camperdown, New South Wales 2006, Australia.
James R
JR
Krycer
Centre for Cancer Biology, SA Pathology and the University of South Australia, Adelaide, South Australia 5000, Australia.
Joanna
J
Woodcock
Heart Research Institute, Thrombosis Group, Newtown, New South Wales 2042, Australia.|Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Jessica
J
Maclean
Centre for Cancer Biology, SA Pathology and the University of South Australia, Adelaide, South Australia 5000, Australia.
Stuart
S
Pitson
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Zhaohua
Z
Zheng
Baker IDI Heart and Diabetes Institute, Melbourne, Victoria 3004, Australia.
Darren C
DC
Henstridge
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Dianne
D
van der Wal
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.|ACRF Department of Cancer Biology and Therapeutics, John Curtin School of Medical Research Australian National University, Canberra ACT 2600, Australia.
Elizabeth E
http://orcid.org/0000-0001-9453-9688
EE
Gardiner
Faculty of Health Sciences, Curtin University, Perth, Western Australia 6102, Australia.
Michael C
MC
Berndt
Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.
Robert K
RK
Andrews
Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|School of Life &Environmental Sciences, The University of Sydney, Camperdown, New South Wales 2006, Australia.|Sydney Medical School, The University of Sydney, Camperdown, New South Wales 2006, Australia.
David E
DE
James
Centre for Cancer Biology, SA Pathology and the University of South Australia, Adelaide, South Australia 5000, Australia.
Angel F
AF
Lopez
Heart Research Institute, Thrombosis Group, Newtown, New South Wales 2042, Australia.|Charles Perkins Centre, Cardiovascular Division, The University of Sydney, Camperdown, New South Wales 2006, Australia.|Australian Centre for Blood Diseases, Alfred Medical Research and Education Precinct (AMREP), Monash University, Prahran, Victoria 3004, Australia.|Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USA.
Shaun P
SP
Jackson
Biochem Biophys Res Commun. 2013 Jun 28;436(2):235-9
23726920
Biochem J. 2004 Apr 15;379(Pt 2):395-408
14744259
J Thromb Haemost. 2011 Oct;9(10):1883-91
21958383
Oncogene. 2015 Mar 19;34(12):1475-86
24727893
Nat Med. 2002 Nov;8(11):1227-34
12411949
Biochemistry. 1998 Jan 13;37(2):638-47
9425086
Blood. 2005 Sep 15;106(6):1975-81
15941906
Oncotarget. 2015 Jun 10;6(16):14522-36
25971334
J Proteome Res. 2008 May;7(5):2078-87
18407684
IUBMB Life. 2002 Jan;53(1):49-55
12018408
Clin Exp Med. 2013 May;13(2):85-97
23456539
Blood. 1998 Feb 15;91(4):1295-303
9454760
Circ Res. 2009 Dec 4;105(12):1177-85
19875727
Biochem Soc Trans. 2002 Aug;30(4):416-21
12196106
Blood. 2004 Nov 15;104(10):3161-8
15271795
Blood. 2000 Jan 15;95(2):551-7
10627461
Transfusion. 2014 Nov;54(11):2911-23
24840017
Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4249-54
11274449
Proc Natl Acad Sci U S A. 2013 Apr 9;110(15):5887-92
23530243
J Thromb Haemost. 2009 Jul;7 Suppl 1:17-20
19630759
J Cell Biol. 1999 Nov 29;147(5):1085-96
10579727
Nat Rev Drug Discov. 2010 Feb;9(2):154-69
20118963
Semin Cell Dev Biol. 2011 Sep;22(7):705-12
21983031
Clin Sci (Lond). 2014 Sep;127(6):367-73
24895057
Blood. 2008 May 1;111(9):4580-7
18299448
J Thromb Haemost. 2010 Feb;8(2):324-30
19874472
Nat Protoc. 2006;1(2):812-23
17406312
Blood. 2009 Jul 16;114(3):663-6
19387006
J Biol Chem. 1994 Jul 15;269(28):18287-90
8034572
Redox Biol. 2014 Jan 10;2:206-10
24494194
Int J Mol Sci. 2012;13(5):5364-74
22754302
Arterioscler Thromb Vasc Biol. 2009 Mar;29(3):316-20
19228604
J Biol Chem. 1996 May 10;271(19):11581-7
8626721
Mol Psychiatry. 2012 Apr;17(4):451-66
22124272
J Thromb Haemost. 2015 Aug;13(8):1372-82
25988873
J Biol Chem. 2003 Aug 29;278(35):33342-50
12810725
Cancer Gene Ther. 2012 Mar;19(3):153-9
22193627
Biochem J. 2004 Jul 15;381(Pt 2):329-42
15167810
Blood. 2010 Sep 23;116(12):2011-8
20538794
Curr Biol. 1994 Apr 1;4(4):315-24
7922340
J Thromb Haemost. 2009 Sep;7(9):1550-5
19558434
FEBS Lett. 2003 May 8;542(1-3):53-9
12729897
J Proteome Res. 2009 Jun;8(6):3006-19
19364129
Blood. 2009 Feb 26;113(9):2056-63
18832659
Cell. 2007 Mar 23;128(6):1173-86
17382885
Blood. 2006 Jun 1;107(11):4346-53
16391010
Blood. 2000 Jul 15;96(2):577-84
10887121
Plant Mol Biol. 2002 Dec;50(6):1041-51
12516871
J Biol Chem. 2004 Jul 30;279(31):32196-204
15166241
Blood. 2004 Jul 15;104(2):420-7
15054037
Semin Cell Dev Biol. 2011 Sep;22(7):713-9
21888985
Trends Cell Biol. 2009 Jan;19(1):16-23
19027299
J Biol Chem. 2000 Jun 2;275(22):16779-87
10828063
J Biochem Mol Toxicol. 2015 May;29(5):240-8
25651404
Int J Oncol. 2004 Aug;25(2):503-9
15254750
Nat Med. 2011 Nov 07;17(11):1423-36
22064432
Cell Calcium. 2011 Sep;50(3):270-8
21616531
J Thromb Haemost. 2006 Sep;4(9):1950-6
16961602
Blood. 2011 Mar 3;117(9):2718-27
21156842
Exp Hematol. 1999 May;27(5):788-96
10340393
Blood. 2013 Jun 13;121(24):4938-47
23584880
Biochim Biophys Acta. 1980 Oct 15;632(3):444-53
7417532
Anal Biochem. 1972 Apr;46(2):489-501
4337047
Blood. 2011 Aug 11;118(6):1663-74
21673344
Blood. 2007 Sep 15;110(6):1879-86
17548580
Cell Cycle. 2014;13(17):2666-70
25486353
Cancer Lett. 2011 Apr 28;303(2):99-107
21334806
J Mol Cell Cardiol. 2015 Jan;78:142-53
25172387
J Biol Chem. 2007 Sep 28;282(39):28648-58
17673465
J Biol Chem. 2002 Feb 22;277(8):6504-10
11724773