Cingulin

Cingulin (CGN; from the Latin cingere “to form a belt around”) is a cytosolic protein encoded by the CGN gene in humans^[5][6][7] localized at tight junctions (TJs) of vertebrate epithelial and endothelial cells.

CGN
Identifiers
Aliases	CGN, cingulin
External IDs	OMIM: 609473 MGI: 1927237 HomoloGene: 41394 GeneCards: CGN
Gene location (Human) Chr. Chromosome 1 (human)^[1] Band 1q21.3 Start 151,510,510 bp^[1] End 151,538,692 bp^[1]
Gene location (Mouse) Chr. Chromosome 3 (mouse)^[2] Band 3 F2.1|3 40.74 cM Start 94,667,376 bp^[2] End 94,693,826 bp^[2]
RNA expression pattern Bgee Human Mouse (ortholog) Top expressed in pancreatic ductal cell right uterine tube body of pancreas rectum right lobe of liver duodenum upper lobe of left lung right lung sural nerve secondary oocyte Top expressed in otolith organ utricle yolk sac jejunum ciliary body ileum secondary oocyte large intestine stomach colon More reference expression data BioGPS n/a
Gene ontology Molecular function actin binding cytoskeletal motor activity protein binding cadherin binding Cellular component myosin complex bicellular tight junction plasma membrane cell junction Biological process transforming growth factor beta receptor signaling pathway biological process Sources:Amigo / QuickGO
Orthologs Species Human Mouse Entrez 57530 70737 Ensembl ENSG00000143375 ENSMUSG00000068876 UniProt Q9P2M7 P59242 RefSeq (mRNA) NM_020770 NM_001037711 NM_001293727 RefSeq (protein) NP_065821 n/a Location (UCSC) Chr 1: 151.51 – 151.54 Mb Chr 3: 94.67 – 94.69 Mb PubMed search ^[3] ^[4]
Wikidata
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Discovery

Cingulin was originally discovered at the MRC Laboratory of Molecular Biology (Cambridge, UK) by Dr. Sandra Citi, as a protein present in chicken intestinal epithelial cells, that co-purified with non-muscle myosin II and was specifically localized at tight junctions (zonulae occludentes).^[8]

Structure & interactions

Cingulin is a homodimer, each subunit containing a N-terminal globular "head" domain, a long α-helical coiled-coil "rod" domain and a small globular C-terminal "tail" region.^[9] This organization is highly conserved throughout vertebrates.^[5] However, cingulin homologs have not been detected in invertebrates.

In vitro, cingulin can bind to and bundle actin filaments, and interact with myosin II and several TJ proteins including ZO-1, ZO-2, ZO-3, paracingulin and occludin.^[10][11][12] Moreover, cingulin forms a complex with JAM-A, a tight junction membrane protein.^[10] Most of cingulin protein interactions are through the globular head domain. Cingulin interacts with ZO-1 through an N-terminal ZO-1 interacting motif (ZIM) in its head region.^[13][14] The rod domain is involved in dimerization and interaction with the RhoA activator, GEF-H1.^[15][16][17]

Cingulin has also been found to interact with microtubules (MTs) through the N-terminal head region, and these interactions was regulated by phosphorylation by the adenosine monophosphate-activated protein kinase (AMPK).^[18]

Function

The function of cingulin has been studied by knockout (KO), knockdown (KD) and over-expression approaches. Embryoid bodies derived from embryonic stem cells where one or both cingulin alleles were targeted by homologous recombination show apparently normal tight junctions, but changes in the expression of a large number of genes, including tight junction protein genes (claudin-2, claudin-6, claudin-7 and occludin) and transcription factors (including GATA4).^[13] Changes in the expression of claudin-2 and ZO-3 are also observed in cultured kidney cells (MDCK) depleted of cingulin by shRNA.^[16]

In 2012, the phenotype of cingulin-knockout mice was described, proving that functional TJ in vivo can be formed in the absence of cingulin.^[19] Together with paracingulin, cingulin also was reported to regulate claudin-2 expression through RhoA-dependent and independent mechanisms.^[19][20] The role of cingulin in development has been studied by morpholino.^[21] oligonucleotide-mediated depletion in chicken, indicating that cingulin is involved in neural crest development. In early mouse and frog embryos, maternal cingulin is localized in the cell cortex. Through early mouse development, cytocortical cingulin in present from oogenesis (cumulus-oocyte contact sites) until 16-cells morulae stage (apical microvillous zones) during early embryogenesis; then maternal cingulin is degraded by endocytic turn-over from the 32-cells stage. Regarding the zygotic cingulin, it accumulates at the tight junctions from 16-cells stage, 10 hours after ZO-1 assembly. Furthermore, the synthesis of cingulin in early mouse embryos is tissue-specific and it occurs in blastocyst (up-regulated in trophectoderm and down-regulated in inner-cells).^[22][23] In Xenopus laevis embryos, maternal cingulin is recruited to apical cell-cell junctions from 2-cells stage.^[24][25]

Homologs

In 2004, a protein homologous to cingulin was discovered and named JACOP (also known as paracingulin, or cingulin-like 1 protein; CGNL1).^[17]

Human diseases

Although cingulin has been involved in regulation of RhoA signaling and gene expression in cultured cells and KO mice, nothing is known about the specific role of cingulin in human diseases.^[15][16][19] Cingulin expression has been studied in human carcinomas and shown to be expressed in adenocarcinomas and down-regulated in squamous carcinomas.^[26][27] Furthermore, histone deacetylase inhibitors, such as sodium butyrate, strongly upregulate its expression in some cultured cells.^[28] Cingulin, as other junctional proteins could be used as a marker of epithelial differentiation, and as a diagnostic marker to distinguish adenocarcinomas from squamous carcinomas.

References

1. GRCh38: Ensembl release 89: ENSG00000143375 - Ensembl, May 2017
2. GRCm38: Ensembl release 89: ENSMUSG00000068876 - Ensembl, May 2017
3. "Human PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
4. "Mouse PubMed Reference:". National Center for Biotechnology Information, U.S. National Library of Medicine.
5. Citi S, D'Atri F, Parry DA (August 2000). "Human and Xenopus cingulin share a modular organization of the coiled-coil rod domain: predictions for intra- and intermolecular assembly". Journal of Structural Biology. 131 (2): 135–45. doi:10.1006/jsbi.2000.4284. PMID 11042084.
6. Nagase T, Kikuno R, Ishikawa KI, Hirosawa M, Ohara O (February 2000). "Prediction of the coding sequences of unidentified human genes. XVI. The complete sequences of 150 new cDNA clones from brain which code for large proteins in vitro". DNA Research. 7 (1): 65–73. doi:10.1093/dnares/7.1.65. PMID 10718198.
7. "Entrez Gene: CGN cingulin".
8. Citi S, Sabanay H, Jakes R, Geiger B, Kendrick-Jones J (May 1988). "Cingulin, a new peripheral component of tight junctions". Nature. 333 (6170): 272–6. Bibcode:1988Natur.333..272C. doi:10.1038/333272a0. PMID 3285223. S2CID 4306432.
9. Cordenonsi M, D'Atri F, Hammar E, Parry DA, Kendrick-Jones J, Shore D, Citi S (December 1999). "Cingulin contains globular and coiled-coil domains and interacts with ZO-1, ZO-2, ZO-3, and myosin". The Journal of Cell Biology. 147 (7): 1569–82. doi:10.1083/jcb.147.7.1569. PMC 2174252. PMID 10613913.
10. Guillemot L, Citi S (2006). "Cingulin, a Cytoskeleton-Associated Protein of the Tight Junction". In Gonzalez-Mariscal L (ed.). Tight junctions. Georgetown, Texas: Landes Bioscience/Eurekah.com. pp. 54–63. ISBN 978-0-387-36673-9.
11. D'Atri F, Citi S (October 2001). "Cingulin interacts with F-actin in vitro". FEBS Letters. 507 (1): 21–4. doi:10.1016/s0014-5793(01)02936-2. PMID 11682052.
12. Cordenonsi M, Turco F, D'atri F, Hammar E, Martinucci G, Meggio F, Citi S (September 1999). "Xenopus laevis occludin. Identification of in vitro phosphorylation sites by protein kinase CK2 and association with cingulin". European Journal of Biochemistry. 264 (2): 374–84. doi:10.1046/j.1432-1327.1999.00616.x. PMID 10491082.
13. Paschoud S, Yu D, Pulimeno P, Jond L, Turner JR, Citi S (February 2011). "Cingulin and paracingulin show similar dynamic behaviour, but are recruited independently to junctions". Molecular Membrane Biology. 28 (2): 123–35. doi:10.3109/09687688.2010.538937. PMC 4336546. PMID 21166484.
14. Pulimeno P, Paschoud S, Citi S (May 2011). "A role for ZO-1 and PLEKHA7 in recruiting paracingulin to tight and adherens junctions of epithelial cells". The Journal of Biological Chemistry. 286 (19): 16743–50. doi:10.1074/jbc.M111.230862. PMC 3089516. PMID 21454477.
15. Citi S, Paschoud S, Pulimeno P, Timolati F, De Robertis F, Jond L, Guillemot L (May 2009). "The tight junction protein cingulin regulates gene expression and RhoA signaling". Annals of the New York Academy of Sciences. 1165 (1): 88–98. Bibcode:2009NYASA1165...88C. doi:10.1111/j.1749-6632.2009.04053.x. PMID 19538293. S2CID 130711.
16. Guillemot L, Citi S (August 2006). "Cingulin regulates claudin-2 expression and cell proliferation through the small GTPase RhoA". Molecular Biology of the Cell. 17 (8): 3569–77. doi:10.1091/mbc.E06-02-0122. PMC 1525245. PMID 16723500.
17. Aijaz S, D'Atri F, Citi S, Balda MS, Matter K (May 2005). "Binding of GEF-H1 to the tight junction-associated adaptor cingulin results in inhibition of Rho signaling and G1/S phase transition". Developmental Cell. 8 (5): 777–86. doi:10.1016/j.devcel.2005.03.003. PMID 15866167.
18. Yano T, Matsui T, Tamura A, Uji M, Tsukita S (November 2013). "The association of microtubules with tight junctions is promoted by cingulin phosphorylation by AMPK". The Journal of Cell Biology. 203 (4): 605–14. doi:10.1083/jcb.201304194. PMC 3840929. PMID 24385485.
19. Guillemot L, Schneider Y, Brun P, Castagliuolo I, Pizzuti D, Martines D, Jond L, Bongiovanni M, Citi S (November 2012). "Cingulin is dispensable for epithelial barrier function and tight junction structure, and plays a role in the control of claudin-2 expression and response to duodenal mucosa injury". Journal of Cell Science. 125 (Pt 21): 5005–14. doi:10.1242/jcs.101261. PMID 22946046.
20. Guillemot L, Spadaro D, Citi S (2013). "The junctional proteins cingulin and paracingulin modulate the expression of tight junction protein genes through GATA-4". PLOS ONE. 8 (2): e55873. Bibcode:2013PLoSO...855873G. doi:10.1371/journal.pone.0055873. PMC 3567034. PMID 23409073.
21. Kos R, Reedy MV, Johnson RL, Erickson CA (April 2001). "The winged-helix transcription factor FoxD3 is important for establishing the neural crest lineage and repressing melanogenesis in avian embryos". Development. 128 (8): 1467–79. doi:10.1242/dev.128.8.1467. PMID 11262245.
22. Javed Q, Fleming TP, Hay M, Citi S (March 1993). "Tight junction protein cingulin is expressed by maternal and embryonic genomes during early mouse development". Development. 117 (3): 1145–51. doi:10.1242/dev.117.3.1145. PMID 8325239.
23. Fleming TP, Hay M, Javed Q, Citi S (March 1993). "Localisation of tight junction protein cingulin is temporally and spatially regulated during early mouse development". Development. 117 (3): 1135–44. doi:10.1242/dev.117.3.1135. PMID 8325238.
24. Cardellini P, Davanzo G, Citi S (September 1996). "Tight junctions in early amphibian development: detection of junctional cingulin from the 2-cell stage and its localization at the boundary of distinct membrane domains in dividing blastomeres in low calcium". Developmental Dynamics. 207 (1): 104–13. doi:10.1002/(SICI)1097-0177(199609)207:1<104::AID-AJA10>3.0.CO;2-0. PMID 8875080. S2CID 23927107.
25. Fesenko I, Kurth T, Sheth B, Fleming TP, Citi S, Hausen P (August 2000). "Tight junction biogenesis in the early Xenopus embryo". Mechanisms of Development. 96 (1): 51–65. doi:10.1016/s0925-4773(00)00368-3. PMID 10940624. S2CID 16765097.
26. Paschoud S, Bongiovanni M, Pache JC, Citi S (September 2007). "Claudin-1 and claudin-5 expression patterns differentiate lung squamous cell carcinomas from adenocarcinomas". Modern Pathology. 20 (9): 947–54. doi:10.1038/modpathol.3800835. PMID 17585317.
27. Citi S, Amorosi A, Franconi F, Giotti A, Zampi G (April 1991). "Cingulin, a specific protein component of tight junctions, is expressed in normal and neoplastic human epithelial tissues". The American Journal of Pathology. 138 (4): 781–9. PMC 1886117. PMID 2012170.
28. Bordin M, D'Atri F, Guillemot L, Citi S (December 2004). "Histone deacetylase inhibitors up-regulate the expression of tight junction proteins". Molecular Cancer Research. 2 (12): 692–701. doi:10.1158/1541-7786.692.2.12. PMID 15634758. S2CID 397815.

Further reading

• Wolburg H, Lippoldt A (June 2002). "Tight junctions of the blood-brain barrier: development, composition and regulation". Vascular Pharmacology. 38 (6): 323–37. doi:10.1016/S1537-1891(02)00200-8. PMID 12529927.
• Bazzoni G, Martinez-Estrada OM, Orsenigo F, Cordenonsi M, Citi S, Dejana E (July 2000). "Interaction of junctional adhesion molecule with the tight junction components ZO-1, cingulin, and occludin". The Journal of Biological Chemistry. 275 (27): 20520–6. doi:10.1074/jbc.M905251199. PMID 10877843.
• D'Atri F, Nadalutti F, Citi S (August 2002). "Evidence for a functional interaction between cingulin and ZO-1 in cultured cells". The Journal of Biological Chemistry. 277 (31): 27757–64. doi:10.1074/jbc.M203717200. PMID 12023291.
• Gevaert K, Goethals M, Martens L, Van Damme J, Staes A, Thomas GR, Vandekerckhove J (May 2003). "Exploring proteomes and analyzing protein processing by mass spectrometric identification of sorted N-terminal peptides". Nature Biotechnology. 21 (5): 566–9. doi:10.1038/nbt810. PMID 12665801. S2CID 23783563.
• Jin J, Smith FD, Stark C, Wells CD, Fawcett JP, Kulkarni S, Metalnikov P, O'Donnell P, Taylor P, Taylor L, Zougman A, Woodgett JR, Langeberg LK, Scott JD, Pawson T (August 2004). "Proteomic, functional, and domain-based analysis of in vivo 14-3-3 binding proteins involved in cytoskeletal regulation and cellular organization". Current Biology. 14 (16): 1436–50. Bibcode:2004CBio...14.1436J. doi:10.1016/j.cub.2004.07.051. PMID 15324660. S2CID 2371325.
• Benzinger A, Muster N, Koch HB, Yates JR, Hermeking H (June 2005). "Targeted proteomic analysis of 14-3-3 sigma, a p53 effector commonly silenced in cancer". Molecular & Cellular Proteomics. 4 (6): 785–95. doi:10.1074/mcp.M500021-MCP200. PMID 15778465.
• Aijaz S, D'Atri F, Citi S, Balda MS, Matter K (May 2005). "Binding of GEF-H1 to the tight junction-associated adaptor cingulin results in inhibition of Rho signaling and G1/S phase transition". Developmental Cell. 8 (5): 777–86. doi:10.1016/j.devcel.2005.03.003. PMID 15866167.
• Kim JE, Tannenbaum SR, White FM (2005). "Global phosphoproteome of HT-29 human colon adenocarcinoma cells". Journal of Proteome Research. 4 (4): 1339–46. doi:10.1021/pr050048h. PMID 16083285.