Protein kinase C fusion proteins are paradoxically loss of function in cancer.
Animals
Binding Sites
COS Cells
Catalytic Domain
Cell Line, Tumor
Chlorocebus aethiops
Diglycerides
/ metabolism
Fluorescence Resonance Energy Transfer
/ methods
Humans
Loss of Function Mutation
/ genetics
Neoplasms
/ metabolism
Phosphorylation
Protein Domains
Protein Kinase C
/ genetics
Protein Kinase C-alpha
/ genetics
Recombinant Fusion Proteins
/ genetics
autoinhibition
cancer
catalytic domain
constitutively active
dominant negative
gene fusions
loss of function
protein degradation
protein kinase C (PKC)
regulatory domain
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
08
11
2020
revised:
11
02
2021
accepted:
18
02
2021
pubmed:
23
2
2021
medline:
31
8
2021
entrez:
22
2
2021
Statut:
ppublish
Résumé
Within the AGC kinase superfamily, gene fusions resulting from chromosomal rearrangements have been most frequently described for protein kinase C (PKC), with gene fragments encoding either the C-terminal catalytic domain or the N-terminal regulatory moiety fused to other genes. Kinase fusions that eliminate regulatory domains are typically gain of function and often oncogenic. However, several quality control pathways prevent accumulation of aberrant PKC, suggesting that PKC fusions may paradoxically be loss of function. To explore this topic, we used biochemical, cellular, and genome editing approaches to investigate the function of fusions that retain the portion of the gene encoding either the catalytic domain or regulatory domain of PKC. Overexpression studies revealed that PKC catalytic domain fusions were constitutively active but vulnerable to degradation. Genome editing of endogenous genes to generate a cancer-associated PKC fusion resulted in cells with detectable levels of fusion transcript but no detectable protein. Hence, PKC catalytic domain fusions are paradoxically loss of function as a result of their instability, preventing appreciable accumulation of protein in cells. Overexpression of a PKC regulatory domain fusion suppressed both basal and agonist-induced endogenous PKC activity, acting in a dominant-negative manner by competing for diacylglycerol. For both catalytic and regulatory domain fusions, the PKC component of the fusion proteins mediated the effects of the full-length fusions on the parameters examined, suggesting that the partner protein is dispensable in these contexts. Taken together, our findings reveal that PKC gene fusions are distinct from oncogenic fusions and present a mechanism by which loss of PKC function occurs in cancer.
Identifiants
pubmed: 33617877
pii: S0021-9258(21)00218-0
doi: 10.1016/j.jbc.2021.100445
pmc: PMC8008189
pii:
doi:
Substances chimiques
1,2-diacylglycerol
0
Diglycerides
0
Recombinant Fusion Proteins
0
Protein Kinase C
EC 2.7.11.13
Protein Kinase C-alpha
EC 2.7.11.13
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
100445Subventions
Organisme : NIGMS NIH HHS
ID : R35 GM122523
Pays : United States
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
Références
Arthritis Rheum. 2013 Aug;65(8):2161-71
pubmed: 23666743
Proc Natl Acad Sci U S A. 1983 Jul;80(14):4208-12
pubmed: 6308606
J Biochem. 2002 Nov;132(5):669-75
pubmed: 12417014
J Neurosci. 2010 Nov 10;30(45):15102-12
pubmed: 21068316
J Cell Biol. 2003 Jun 9;161(5):899-909
pubmed: 12782683
Sci Rep. 2018 Jan 15;8(1):720
pubmed: 29335433
Int J Biochem Cell Biol. 2014 Aug;53:475-81
pubmed: 24721208
Biochemistry. 2018 Aug 7;57(31):4717-4725
pubmed: 29920078
Cell Rep. 2018 Apr 3;23(1):227-238.e3
pubmed: 29617662
Trends Pharmacol Sci. 2017 May;38(5):438-447
pubmed: 28283201
Am J Physiol Lung Cell Mol Physiol. 2000 Sep;279(3):L429-38
pubmed: 10956616
Nat Rev Cancer. 2007 Apr;7(4):281-94
pubmed: 17384583
Genome Res. 2012 Nov;22(11):2109-19
pubmed: 22975805
Cold Spring Harb Perspect Biol. 2019 Sep 3;11(9):
pubmed: 30745292
Cancer Discov. 2018 Mar;8(3):336-353
pubmed: 29242214
Nucleic Acids Res. 2019 Jan 8;47(D1):D941-D947
pubmed: 30371878
Oncogene. 1999 Sep 30;18(40):5604-9
pubmed: 10523838
Blood. 2013 Apr 18;121(16):3117-25
pubmed: 23430113
Lab Invest. 2015 Sep;95(9):1071-6
pubmed: 26121314
J Clin Neurosci. 2016 Jan;23:73-75
pubmed: 26260115
EMBO J. 1999 Nov 15;18(22):6329-38
pubmed: 10562545
Proc Natl Acad Sci U S A. 2009 Apr 21;106(16):6603-7
pubmed: 19346474
J Biol Chem. 2007 Nov 16;282(46):33776-33787
pubmed: 17893151
Nucleic Acids Res. 2018 Jan 4;46(D1):D1144-D1149
pubmed: 29099951
Nature. 2017 Aug 17;548(7667):297-303
pubmed: 28783718
Anal Biochem. 1976 May 7;72:248-54
pubmed: 942051
Exp Biol Med (Maywood). 2019 May;244(8):655-662
pubmed: 30776907
Kidney Int Suppl. 2007 Aug;(106):S49-53
pubmed: 17653211
Blood. 2013 Apr 18;121(16):3112-6
pubmed: 23319571
Curr Biol. 1995 Dec 1;5(12):1394-1403
pubmed: 8749392
Oncotarget. 2018 Sep 28;9(76):34306-34319
pubmed: 30344944
Nat Genet. 2017 Aug;49(8):1211-1218
pubmed: 28671688
Elife. 2018 Jul 03;7:
pubmed: 29968564
Mol Cell. 2019 Apr 18;74(2):378-392.e5
pubmed: 30904392
Sci Rep. 2017 Jul 28;7(1):6829
pubmed: 28754924
J Biol Chem. 2007 Jan 12;282(2):826-30
pubmed: 17071619
Oncotarget. 2016 Apr 12;7(15):20919-33
pubmed: 26989024
Chem Biol. 2014 Apr 24;21(4):459-469
pubmed: 24631122
J Biol Chem. 1998 Apr 10;273(15):8669-74
pubmed: 9535842
Methods Mol Biol. 2013;946:1-13
pubmed: 23179822
J Biol Chem. 2006 Oct 13;281(41):30947-56
pubmed: 16901905
J Biol Chem. 2003 Sep 5;278(36):33753-62
pubmed: 12824193
Brain Pathol. 2013 Mar;23(2):121-8
pubmed: 22725730
Cell Death Differ. 1999 Feb;6(2):99-104
pubmed: 10200555
Curr Biol. 1999 Jul 15;9(14):728-37
pubmed: 10421574
Nucleic Acids Res. 2015 Apr 20;43(7):e45
pubmed: 25586224
Biochim Biophys Acta Mol Cell Res. 2018 Jun;1865(6):889-897
pubmed: 29563055
Neuronal Signal. 2017 Apr 10;1(2):NS20160005
pubmed: 32714576
Nat Rev Cancer. 2015 Jun;15(6):371-81
pubmed: 25998716
J Biol Chem. 1982 Jul 10;257(13):7847-51
pubmed: 7085651
J Thorac Oncol. 2019 Dec;14(12):e266-e268
pubmed: 31757376
Nat Commun. 2014 Sep 10;5:4846
pubmed: 25204415
Haematologica. 2020 May;105(5):1262-1273
pubmed: 31439673
Biochem J. 2021 Jan 29;478(2):341-355
pubmed: 33502516
Proc Natl Acad Sci U S A. 2017 Dec 12;114(50):13076-13084
pubmed: 29162699
Genes Chromosomes Cancer. 2015 Nov;54(11):681-91
pubmed: 26227178
J Biol Chem. 2008 Mar 7;283(10):6300-11
pubmed: 18162466
Mol Cell Biol. 2015 Jan;35(1):264-76
pubmed: 25348715
Genes Cancer. 2012 May;3(5-6):436-46
pubmed: 23226581
Acta Neuropathol Commun. 2015 Dec 15;3:85
pubmed: 26671581
J Biol Chem. 2016 Jun 24;291(26):13809-22
pubmed: 27143478
Cell. 2015 Jan 29;160(3):489-502
pubmed: 25619690
J Biol Chem. 1982 Nov 25;257(22):13193-6
pubmed: 7142138
Nucleic Acid Ther. 2015 Dec;25(6):287-96
pubmed: 26540648
PLoS One. 2015 Jun 08;10(6):e0129047
pubmed: 26053850
Nucleic Acids Res. 2017 Jan 4;45(D1):D784-D789
pubmed: 27899563
Biochemistry. 2000 Sep 19;39(37):11360-9
pubmed: 10985781
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
Trends Biochem Sci. 2011 Feb;36(2):65-77
pubmed: 20971646
Cytokine Growth Factor Rev. 2020 Apr;52:56-67
pubmed: 31899106
Front Oncol. 2020 Nov 25;10:590755
pubmed: 33324562
J Biol Chem. 1994 Jan 21;269(3):2118-24
pubmed: 8294465
Mol Ther. 2019 Jan 2;27(1):200-218
pubmed: 30509566
Biochemistry. 2003 Sep 30;42(38):11194-202
pubmed: 14503869
J Biol Chem. 1983 Oct 10;258(19):11442-5
pubmed: 6311812
Oncogene. 2014 Nov 6;33(45):5225-37
pubmed: 24336328
Int J Mol Sci. 2018 Feb 07;19(2):
pubmed: 29414922
Am J Physiol Endocrinol Metab. 2010 Mar;298(3):E395-402
pubmed: 19934406
Science. 1998 Aug 28;281(5381):1312-6
pubmed: 9721091
EMBO Rep. 2001 Dec;2(12):1133-8
pubmed: 11743028
Curr Biol. 1998 Dec 17-31;8(25):1366-75
pubmed: 9889098
Nat Rev Cancer. 2007 Apr;7(4):233-45
pubmed: 17361217
J Biol Chem. 2009 Feb 20;284(8):4921-35
pubmed: 19091746
Brain Struct Funct. 2019 Jan;224(1):277-292
pubmed: 30315415
Cancer Genet. 2015 Nov;208(11):545-51
pubmed: 26432191
Oncogene. 2015 Sep 10;34(37):4845-54
pubmed: 25500544