Unraveling the hidden role of a uORF-encoded peptide as a kinase inhibitor of PKCs.
PKC
kinase inhibitor
pseudosubstrate
uORF
Journal
Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876
Informations de publication
Date de publication:
05 10 2021
05 10 2021
Historique:
accepted:
19
08
2021
entrez:
1
10
2021
pubmed:
2
10
2021
medline:
15
12
2021
Statut:
ppublish
Résumé
Approximately 40% of human messenger RNAs (mRNAs) contain upstream open reading frames (uORFs) in their 5' untranslated regions. Some of these uORF sequences, thought to attenuate scanning ribosomes or lead to mRNA degradation, were recently shown to be translated, although the function of the encoded peptides remains unknown. Here, we show a uORF-encoded peptide that exhibits kinase inhibitory functions. This uORF, upstream of the protein kinase C-eta (PKC-η) main ORF, encodes a peptide (uPEP2) containing the typical PKC pseudosubstrate motif present in all PKCs that autoinhibits their kinase activity. We show that uPEP2 directly binds to and selectively inhibits the catalytic activity of novel PKCs but not of classical or atypical PKCs. The endogenous deletion of uORF2 or its overexpression in MCF-7 cells revealed that the endogenously translated uPEP2 reduces the protein levels of PKC-η and other novel PKCs and restricts cell proliferation. Functionally, treatment of breast cancer cells with uPEP2 diminished cell survival and their migration and synergized with chemotherapy by interfering with the response to DNA damage. Furthermore, in a xenograft of MDA-MB-231 breast cancer tumor in mice models, uPEP2 suppressed tumor progression, invasion, and metastasis. Tumor histology showed reduced proliferation, enhanced cell death, and lower protein expression levels of novel PKCs along with diminished phosphorylation of PKC substrates. Hence, our study demonstrates that uORFs may encode biologically active peptides beyond their role as translation regulators of their downstream ORFs. Together, we point to a unique function of a uORF-encoded peptide as a kinase inhibitor, pertinent to cancer therapy.
Identifiants
pubmed: 34593629
pii: 2018899118
doi: 10.1073/pnas.2018899118
pmc: PMC8501901
pii:
doi:
Substances chimiques
Peptides
0
Protein Kinase Inhibitors
0
Protein Kinase C
EC 2.7.11.13
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2021 the Author(s). Published by PNAS.
Déclaration de conflit d'intérêts
The authors declare no competing interest.
Références
Proteomics. 2018 May;18(10):e1700038
pubmed: 28759167
J Biol Chem. 2003 Apr 18;278(16):14555-64
pubmed: 12566450
Science. 2009 Apr 10;324(5924):218-23
pubmed: 19213877
Elife. 2015 Dec 19;4:e08890
pubmed: 26687005
Genome Res. 2015 Dec;25(12):1836-47
pubmed: 26338483
Elife. 2015 Feb 26;4:
pubmed: 25719440
Biochem J. 1998 Jun 1;332 ( Pt 2):281-92
pubmed: 9601053
Biochem Biophys Res Commun. 2011 Aug 26;412(2):313-7
pubmed: 21820409
Semin Cancer Biol. 2018 Feb;48:36-52
pubmed: 28571764
Cold Spring Harb Perspect Biol. 2019 May 1;11(5):
pubmed: 30037969
Sci Signal. 2011 Dec 06;4(202):ra84
pubmed: 22155788
J Mol Cell Cardiol. 2011 Nov;51(5):665-73
pubmed: 21810427
Crit Rev Biochem Mol Biol. 2018 Apr;53(2):208-230
pubmed: 29513138
Curr Opin Cell Biol. 1997 Apr;9(2):161-7
pubmed: 9069266
Cancer Lett. 2007 Feb 8;246(1-2):173-81
pubmed: 16580129
Biochem J. 2003 Mar 1;370(Pt 2):361-71
pubmed: 12495431
Proc Natl Acad Sci U S A. 2014 Jan 7;111(1):E203-12
pubmed: 24367078
Science. 2014 Jan 3;343(6166):84-87
pubmed: 24336571
Electrophoresis. 1997 Dec;18(15):2714-23
pubmed: 9504803
Genome Res. 2012 Nov;22(11):2208-18
pubmed: 22879431
Biochem Soc Trans. 2014 Dec;42(6):1519-23
pubmed: 25399563
Oncogene. 2001 Oct 11;20(46):6794-804
pubmed: 11709714
Cell Death Dis. 2014 Nov 20;5:e1531
pubmed: 25412309
J Biol Chem. 2016 Aug 12;291(33):16927-35
pubmed: 27358398
Front Plant Sci. 2012 Aug 24;3:191
pubmed: 22936940
Nat Rev Drug Discov. 2012 Dec;11(12):937-57
pubmed: 23197040
Biochem Biophys Res Commun. 2000 Dec 9;279(1):103-7
pubmed: 11112424
Cancers (Basel). 2019 Feb 13;11(2):
pubmed: 30781807
Exp Cell Res. 2011 Jun 10;317(10):1465-75
pubmed: 21514295
Science. 1987 Dec 18;238(4834):1726-8
pubmed: 3686012
Mol Cell. 2010 Oct 22;40(2):228-37
pubmed: 20965418
Cell. 2011 Nov 11;147(4):789-802
pubmed: 22056041
Neuro Oncol. 2002 Jan;4(1):9-21
pubmed: 11772428
Onkologie. 2012;35(5):260-6
pubmed: 22868505
J Biol Chem. 1997 Jan 10;272(2):952-60
pubmed: 8995387
Chem Rev. 2001 Aug;101(8):2353-64
pubmed: 11749377
PLoS Genet. 2013;9(8):e1003529
pubmed: 23950723
EMBO J. 2016 Apr 1;35(7):706-23
pubmed: 26896445
Mol Cancer Ther. 2004 Jul;3(7):773-81
pubmed: 15252138
Cell. 1987 Jun 19;49(6):805-13
pubmed: 3555844
Nature. 2013 Jan 17;493(7432):420-3
pubmed: 23283174
J Biol Chem. 2001 Dec 21;276(51):47759-62
pubmed: 11673449
Elife. 2017 Oct 30;6:
pubmed: 29083303
J Biol Chem. 2001 Oct 12;276(41):38036-43
pubmed: 11489903
Nucleic Acids Res. 2014 Jan;42(Database issue):D859-64
pubmed: 24185699
Mol Cell Biol. 2000 Dec;20(23):8635-42
pubmed: 11073965
Genome Res. 2015 Nov;25(11):1610-21
pubmed: 26297486
Cancer Biol Ther. 2007 Sep;6(9):1375-80
pubmed: 17786031
Science. 2020 Mar 6;367(6482):1140-1146
pubmed: 32139545
Mol Carcinog. 1994 Dec;11(4):197-203
pubmed: 7999261
Nat Rev Mol Cell Biol. 2010 Feb;11(2):103-12
pubmed: 20094051
PLoS One. 2012;7(10):e47995
pubmed: 23118918
Exp Cell Res. 2009 Sep 10;315(15):2616-23
pubmed: 19523467
Proc Natl Acad Sci U S A. 2012 Sep 11;109(37):E2424-32
pubmed: 22927429
Nat Methods. 2014 Aug;11(8):783-784
pubmed: 25075903
J Biol Chem. 1998 Nov 13;273(46):30306-15
pubmed: 9804792
Mol Cell Biol. 2009 Nov;29(22):6140-8
pubmed: 19797084
J Biol Chem. 2014 Jan 17;289(3):1257-70
pubmed: 24293369
Cell. 2015 Jan 29;160(3):489-502
pubmed: 25619690
Am J Physiol Endocrinol Metab. 2010 Mar;298(3):E395-402
pubmed: 19934406
Am J Physiol Regul Integr Comp Physiol. 2014 Apr 15;306(8):R619-26
pubmed: 24523339
EMBO Rep. 2015 Dec;16(12):1723-36
pubmed: 26538417
Physiol Rev. 2008 Oct;88(4):1341-78
pubmed: 18923184
Mol Cell. 2015 Dec 3;60(5):816-827
pubmed: 26638175
Cancer Res. 2003 Aug 1;63(15):4347-50
pubmed: 12907603