A Novel Conserved Domain Mediates Dimerization of Protein Kinase D (PKD) Isoforms: DIMERIZATION IS ESSENTIAL FOR PKD-DEPENDENT REGULATION OF SECRETION AND INNATE IMMUNITY.
Protein kinase D (PKD) isoforms are protein kinase C effectors in signaling pathways regulated by diacylglycerol. Important physiological processes (including secretion, immune responses, motility, and transcription) are placed under diacylglycerol control by the distinctive substrate specificity and subcellular distribution of PKDs. Potentially, broadly co-expressed PKD polypeptides may interact to generate homo- or heteromultimeric regulatory complexes. However, the frequency, molecular basis, regulatory significance, and physiological relevance of stable PKD-PKD interactions are largely unknown. Here, we demonstrate that mammalian PKDs 1-3 and the prototypical Caenorhabditis elegans PKD, DKF-2A, are exclusively (homo- or hetero-) dimers in cell extracts and intact cells. We discovered and characterized a novel, highly conserved N-terminal domain, comprising 92 amino acids, which mediates dimerization of PKD1, PKD2, and PKD3 monomers. A similar domain directs DKF-2A homodimerization. Dimerization occurred independently of properties of the regulatory and kinase domains of PKDs. Disruption of PKD dimerization abrogates secretion of PAUF, a protein carried in small trans-Golgi network-derived vesicles. In addition, disruption of DKF-2A homodimerization in C. elegans intestine impaired and degraded the immune defense of the intact animal against an ingested bacterial pathogen. Finally, dimerization was indispensable for the strong, dominant negative effect of catalytically inactive PKDs. Overall, the structural integrity and function of the novel dimerization domain are essential for PKD-mediated regulation of a key aspect of cell physiology, secretion, and innate immunity in vivo.
From the Department of Molecular Pharmacology, Atran Laboratories, Albert Einstein College of Medicine, Bronx, New York 10461.
Clara
http://orcid.org/0000-0002-1114-4259
C
Aicart-Ramos
From the Department of Molecular Pharmacology, Atran Laboratories, Albert Einstein College of Medicine, Bronx, New York 10461.
Sophia Dan Qing
SD
He
From the Department of Molecular Pharmacology, Atran Laboratories, Albert Einstein College of Medicine, Bronx, New York 10461.
Marianne
M
Land
From the Department of Molecular Pharmacology, Atran Laboratories, Albert Einstein College of Medicine, Bronx, New York 10461 charles.rubin@einstein.yu.edu.
Charles S
CS
Rubin
D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000737:chemistry / Q000276:immunology; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry / Q000276:immunology; D017124:Conserved Sequence; D057809:HEK293 Cells; D006801:Humans; D007113:Immunity, Innate; D000072417:Protein Domains; D020033:Protein Isoforms / Q000737:chemistry / Q000276:immunology; D011493:Protein Kinase C / Q000737:chemistry* / Q000276:immunology; D055503:Protein Multimerization; D016415:Sequence Alignment
D029742:Caenorhabditis elegans Proteins; D020033:Protein Isoforms; C524695:DKF-2 protein, C elegans; C089212:protein kinase D; D011493:Protein Kinase C
Caenorhabditis elegans (C. elegans); heterodimer; homodimer; innate immunity; native Mr; oligomerization domain; protein kinase D (PKD); secretion; signal transduction
Dev Cell. 2012 Oct 16;23(4):756-68
22981988
Am J Physiol Endocrinol Metab. 2010 Mar;298(3):E395-402
19934406
PLoS One. 2014 May 19;9(5):e98090
24840177
IUBMB Life. 2013 Feb;65(2):98-107
23288632
J Exp Med. 2005 Mar 7;201(5):793-804
15738054
J Biol Chem. 1994 Mar 25;269(12):9234-44
8132661
Sci Signal. 2009 Aug 11;2(83):ra42
19671928
Bioarchitecture. 2014;4(3):111-5
24847910
Annu Rev Biochem. 2012;81:407-27
22463692
J Biol Chem. 2001 Aug 31;276(35):32616-26
11410587
J Biol Chem. 1998 Jan 9;273(2):1130-43
9422779
J Biol Chem. 2007 Oct 26;282(43):31273-88
17728253
Cell Signal. 2008 May;20(5):925-34
18262756
Nat Cell Biol. 2004 Feb;6(2):106-12
14743217
Oncogene. 2011 Jan 13;30(2):201-11
20802527
Physiol Rev. 2008 Oct;88(4):1341-78
18923184
Mol Cancer Ther. 2015 Jun;14(6):1306-16
25852060
Sci Signal. 2014 Oct 21;7(348):ra99
25336615
Cell Mol Life Sci. 2015 Nov;72 (22):4369-82
26253275
Expert Rev Gastroenterol Hepatol. 2015;9(9):1143-6
26174103
J Clin Med. 2016 Feb 03;5(2):null
26848698
Mol Biol Cell. 1992 Nov;3(11):1215-28
1333841
J Med Chem. 2010 Aug 12;53(15):5422-38
20684592
Mol Cell Biol. 2008 Jun;28(11):3600-9
18378694
Immunity. 2009 Apr 17;30(4):521-32
19371715
Biochim Biophys Acta. 1966 Feb 7;112(2):346-62
5329026
Oncogene. 2013 Aug 1;32(31):3638-47
22907431
Nat Cell Biol. 2009 May;11(5):545-56
19329994
J Cell Sci. 2012 Oct 15;125(Pt 20):4800-11
22797919
J Cell Biol. 2002 Jan 7;156(1):65-74
11777941
EMBO Rep. 2011 Jul 08;12(8):785-96
21738220
Cell Signal. 2013 Sep;25(9):1739-47
23688773
J Biol Chem. 2008 Feb 1;283(5):2614-21
18048355
Biochem J. 2012 Mar 15;442(3):649-59
22233340
EMBO J. 2012 Oct 17;31(20):3976-90
22909819
Mol Cancer Ther. 2010 May;9(5):1136-46
20442301
Mol Cell Biol. 2006 Feb;26(4):1569-77
16449666
Cell. 1995 Jun 16;81(6):917-24
7781068
Biochem J. 2010 Nov 15;432(1):153-63
20819079
Nat Cell Biol. 2005 Sep;7(9):880-6
16100512
J Biol Chem. 1997 Jan 10;272(2):952-60
8995387
Gene. 1990 Sep 14;93(2):189-98
2121610
J Cell Biol. 2007 Dec 17;179(6):1123-31
18086912
Cell. 1998 Oct 30;95(3):307-18
9814702
Cancer Sci. 2009 May;100(5):828-36
19302292
Biol Proced Online. 2009 May 15;11:32-51
19495910
J Biol Chem. 1989 Mar 15;264(8):4648-56
2538452
Trends Cell Biol. 2007 Jan;17(1):13-8
17126550
J Biol Chem. 1993 Apr 15;268(11):7621-4
8463292
Proc Natl Acad Sci U S A. 2008 Feb 26;105(8):3059-63
18287012
Cardiovasc Res. 2007 Mar 1;73(4):667-77
17217938
Physiology (Bethesda). 2011 Feb;26(1):23-33
21357900
J Biol Chem. 2006 Jun 30;281(26):17801-14
16613841
PLoS One. 2013 Sep 23;8(9):e75601
24086585