Cingulin binds to the ZU5 domain of scaffolding protein ZO-1 to promote its extended conformation, stabilization, and tight junction accumulation.
ZO-1
cingulin
paracingulin
tight junctions
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:
04 2022
04 2022
Historique:
received:
15
10
2021
revised:
21
02
2022
accepted:
23
02
2022
pubmed:
9
3
2022
medline:
27
4
2022
entrez:
8
3
2022
Statut:
ppublish
Résumé
Zonula occludens-1 (ZO-1), the major scaffolding protein of tight junctions (TJs), recruits the cytoskeleton-associated proteins cingulin (CGN) and paracingulin (CGNL1) to TJs by binding to their N-terminal ZO-1 interaction motif. The conformation of ZO-1 can be either folded or extended, depending on cytoskeletal tension and intramolecular and intermolecular interactions, and only ZO-1 in the extended conformation recruits the transcription factor DbpA to TJs. However, the sequences of ZO-1 that interact with CGN and CGNL1 and the role of TJ proteins in ZO-1 TJ assembly are not known. Here, we used glutathione-S-transferase pulldowns and immunofluorescence microscopy to show that CGN and CGNL1 bind to the C-terminal ZU5 domain of ZO-1 and that this domain is required for CGN and CGNL1 recruitment to TJs and to phase-separated ZO-1 condensates in cells. We show that KO of CGN, but not CGNL1, results in decreased accumulation of ZO-1 at TJs. Furthermore, ZO-1 lacking the ZU5 domain showed decreased accumulation at TJs, was detectable along lateral contacts, had a higher mobile fraction than full-length ZO-1 by fluorescence recovery after photobleaching analysis, and had a folded conformation, as determined by structured illumination microscopy of its N-terminal and C-terminal ends. The CGN-ZU5 interaction promotes the extended conformation of ZO-1, since binding of the CGN-ZO-1 interaction motif region to ZO-1 resulted in its interaction with DbpA in cells and in vitro. Together, these results show that binding of CGN to the ZU5 domain of ZO-1 promotes ZO-1 stabilization and accumulation at TJs by promoting its extended conformation.
Identifiants
pubmed: 35259394
pii: S0021-9258(22)00237-X
doi: 10.1016/j.jbc.2022.101797
pmc: PMC9010756
pii:
doi:
Substances chimiques
Cytoskeletal Proteins
0
Phosphoproteins
0
Zonula Occludens-1 Protein
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
101797Subventions
Organisme : NIDDK NIH HHS
ID : R01 DK061931
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK068271
Pays : United States
Informations de copyright
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest J. R. T. is a founder and shareholder of Thelium Therapeutics and has served as a consultant for Entrinsic, Immunic, and Kallyope. S. C. has served as a consultant for Immunic. All other authors declare that they have no conflicts of interest with the contents of this article.
Références
Tissue Barriers. 2019;7(3):e1653748
pubmed: 31438766
J Cell Biol. 1993 Oct;123(2):293-302
pubmed: 8408213
J Biol Chem. 1998 Nov 6;273(45):29745-53
pubmed: 9792688
Nat Cell Biol. 2011 Feb;13(2):159-66
pubmed: 21258369
J Biol Chem. 2004 Oct 22;279(43):44785-94
pubmed: 15292177
Cell. 2019 Oct 31;179(4):937-952.e18
pubmed: 31675500
Cell. 2019 Oct 31;179(4):923-936.e11
pubmed: 31675499
Trends Biochem Sci. 2019 Feb;44(2):141-152
pubmed: 30665499
Ann N Y Acad Sci. 2009 May;1165:113-20
pubmed: 19538295
Tissue Barriers. 2018;6(3):1539596
pubmed: 30395792
Mol Biol Cell. 2021 Nov 1;32(21):ar34
pubmed: 34613798
Ann N Y Acad Sci. 2012 Jun;1257:125-32
pubmed: 22671598
Nat Commun. 2016 Aug 03;7:12426
pubmed: 27484926
J Biol Chem. 2016 May 20;291(21):11016-29
pubmed: 27044745
Trends Cell Biol. 2020 Oct;30(10):805-817
pubmed: 32891490
J Cell Biol. 2000 Nov 13;151(4):825-36
pubmed: 11076967
J Biol Chem. 2014 Aug 8;289(32):22500-11
pubmed: 24986862
Mol Biol Cell. 2009 Sep;20(17):3930-40
pubmed: 19605556
Nature. 1988 May 19;333(6170):272-6
pubmed: 3285223
J Am Soc Nephrol. 2014 Feb;25(2):250-9
pubmed: 24179170
Proc Natl Acad Sci U S A. 2012 Jun 19;109(25):9905-10
pubmed: 22665792
J Biol Chem. 2011 May 13;286(19):16743-50
pubmed: 21454477
Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3460-4
pubmed: 2014265
Curr Biol. 2017 Dec 18;27(24):3783-3795.e8
pubmed: 29199076
Proc Natl Acad Sci U S A. 2010 May 4;107(18):8237-41
pubmed: 20404178
J Biol Chem. 2012 Apr 13;287(16):13159-69
pubmed: 22315225
PLoS One. 2010 Aug 20;5(8):e12207
pubmed: 20808826
FEBS Lett. 2001 Oct 19;507(1):21-4
pubmed: 11682052
Sci Rep. 2018 Oct 19;8(1):15550
pubmed: 30341325
J Biol Chem. 2004 Oct 29;279(44):46014-22
pubmed: 15292197
J Cell Biol. 1986 Sep;103(3):755-66
pubmed: 3528172
Cell. 2006 Aug 25;126(4):741-54
pubmed: 16923393
EMBO Rep. 2009 Oct;10(10):1125-31
pubmed: 19730435
J Biol Chem. 2018 Nov 9;293(45):17317-17335
pubmed: 30242130
J Cell Biol. 1996 Mar;132(6):1115-32
pubmed: 8601589
Ann N Y Acad Sci. 2017 Oct;1405(1):160-176
pubmed: 28617990
J Mol Med (Berl). 2014 Mar;92(3):217-25
pubmed: 24553779
Nat Rev Mol Cell Biol. 2016 Sep;17(9):564-80
pubmed: 27353478
Mol Biol Cell. 2006 Apr;17(4):1922-32
pubmed: 16436508
J Biol Chem. 2013 May 10;288(19):13775-88
pubmed: 23553632
Dev Dyn. 1996 Sep;207(1):104-13
pubmed: 8875080
J Cell Biol. 1999 Dec 27;147(7):1569-82
pubmed: 10613913
J Cell Biol. 2003 Feb 3;160(3):423-32
pubmed: 12566432
J Biol Chem. 1999 Dec 3;274(49):35179-85
pubmed: 10575001
Cold Spring Harb Perspect Biol. 2009 Aug;1(2):a002584
pubmed: 20066090
Curr Biol. 2020 Jan 20;30(2):R83-R86
pubmed: 31962084
J Cell Biol. 2008 May 19;181(4):683-95
pubmed: 18474622
J Cell Sci. 2013 Sep 1;126(Pt 17):3904-15
pubmed: 23813953
Cell Rep. 2018 Nov 20;25(8):2132-2147.e7
pubmed: 30463011
Mol Biol Cell. 2008 Jun;19(6):2465-75
pubmed: 18353970
EMBO J. 2011 Feb 16;30(4):665-78
pubmed: 21240187
J Biol Chem. 2002 Aug 2;277(31):27757-64
pubmed: 12023291
Mech Dev. 2000 Aug;96(1):51-65
pubmed: 10940624
Biochim Biophys Acta Biomembr. 2020 Oct 1;1862(10):183399
pubmed: 32553946
Prog Biophys Mol Biol. 2003 Jan;81(1):1-44
pubmed: 12475568
Tissue Barriers. 2014 Jan 1;2(1):e28755
pubmed: 24843844