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Protocadherin-1 binds to SMAD3 and suppresses TGF-?1-induced gene transcription.

Protocadherin-1 binds to SMAD3 and suppresses TGF-?1-induced gene transcription.
Protocadherin-1 binds to SMAD3 and suppresses TGF-?1-induced gene transcription.
Genetic studies have identified Protocadherin-1 (PCDH1) and Mothers against decapentaplegic homolog-3 (SMAD3) as susceptibility genes for asthma. PCDH1 is expressed in bronchial epithelial cells and has been found to interact with SMAD3 in yeast two-hybrid (Y2H) overexpression assays. Here, we test whether PCDH1 and SMAD3 interact at endogenous protein levels in bronchial epithelial cells and evaluate the consequences thereof for transforming growth factor-?1 (TGF-?1)-induced gene transcription. We performed Y2H screens and coimmunoprecipitation (co-IP) experiments of PCDH1 and SMAD3 in HEK293T and 16HBE14o(-) (16HBE) cell lines. Activity of a SMAD3-driven luciferase reporter gene in response to TGF-?1 was measured in BEAS-2B cells transfected with PCDH1 and in 16HBE cells transfected with PCDH1-small-interfering RNA (siRNA). TGF-?1-induced gene expression was quantified in BEAS-2B clones overexpressing PCDH1 and in human primary bronchial epithelial cells (PBECs) transfected with PCDH1-siRNA. We confirm PCDH1 and SMAD3 interactions by Y2H and by co-IP in HEK293T cells overexpressing both proteins, and at endogenous protein levels in 16HBE cells. TGF-?-induced activation of a SMAD3-driven reporter was reduced by exogenous PCDH1 in BEAS2B cells, whereas it was increased by siRNA-mediated knockdown of endogenous PCDH1 in 16HBE cells. Overexpression of PCDH1 suppressed expression of TGF-? target genes in BEAS-2B cells, whereas knockdown of PCDH1 in human PBECs increased TGF-?-induced gene expression. In conclusion, we demonstrate that PCDH1 binds to SMAD3 and regulates its activation by TGF-? signaling in bronchial epithelial cells. We propose that PCDH1 and SMAD3 act in a single pathway in asthma susceptibility that affects sensitivity of the airway epithelium to TGF-?.
Mothers against decapentaplegic homolog-3, airway epithelium, functional genetics, asthma susceptibility, transforming growth factor-?-induced gene expression
1040-0605
L725-L735
Faura Tellez, Grissel
a4dcdc41-af1d-444a-b6c4-14f4efd1a93c
Vandepoele, Karl
56deacff-22d8-469d-b3ab-d4a659a943aa
Brouwer, Uilke
260b1582-747d-48bf-9ca5-a16bdd9fda56
Koning, Henk
f053a060-7e1c-4012-b21e-acd9883280ef
Elderman, Robin M.
df50c4be-37f9-48b3-ac3f-538ed5ec9229
Hackett, Tillie-Louise
05158d38-5bd8-4cce-9c82-5f86ab8f2757
Willemse, Brigitte W.M.
bf59c206-763b-40e1-b5de-692e80506cf2
Holloway, John
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Van Roy, Frans
0c236c08-cfcb-4231-838b-d42162757f93
Koppelman, Gerard H.
8d04aab8-5795-4e62-b64a-de7e368c0d1d
Nawijn, Martijn C.
16a331eb-87b5-48e7-976f-ddbb3fc9efe9
Faura Tellez, Grissel
a4dcdc41-af1d-444a-b6c4-14f4efd1a93c
Vandepoele, Karl
56deacff-22d8-469d-b3ab-d4a659a943aa
Brouwer, Uilke
260b1582-747d-48bf-9ca5-a16bdd9fda56
Koning, Henk
f053a060-7e1c-4012-b21e-acd9883280ef
Elderman, Robin M.
df50c4be-37f9-48b3-ac3f-538ed5ec9229
Hackett, Tillie-Louise
05158d38-5bd8-4cce-9c82-5f86ab8f2757
Willemse, Brigitte W.M.
bf59c206-763b-40e1-b5de-692e80506cf2
Holloway, John
4bbd77e6-c095-445d-a36b-a50a72f6fe1a
Van Roy, Frans
0c236c08-cfcb-4231-838b-d42162757f93
Koppelman, Gerard H.
8d04aab8-5795-4e62-b64a-de7e368c0d1d
Nawijn, Martijn C.
16a331eb-87b5-48e7-976f-ddbb3fc9efe9

Faura Tellez, Grissel, Vandepoele, Karl, Brouwer, Uilke, Koning, Henk, Elderman, Robin M., Hackett, Tillie-Louise, Willemse, Brigitte W.M., Holloway, John, Van Roy, Frans, Koppelman, Gerard H. and Nawijn, Martijn C. (2015) Protocadherin-1 binds to SMAD3 and suppresses TGF-?1-induced gene transcription. American Journal of Physiology: Lung Cellular and Molecular Physiology, 309 (7), L725-L735. (doi:10.1152/ajplung.00346.2014). (PMID:26209277)

Record type: Article

Abstract

Genetic studies have identified Protocadherin-1 (PCDH1) and Mothers against decapentaplegic homolog-3 (SMAD3) as susceptibility genes for asthma. PCDH1 is expressed in bronchial epithelial cells and has been found to interact with SMAD3 in yeast two-hybrid (Y2H) overexpression assays. Here, we test whether PCDH1 and SMAD3 interact at endogenous protein levels in bronchial epithelial cells and evaluate the consequences thereof for transforming growth factor-?1 (TGF-?1)-induced gene transcription. We performed Y2H screens and coimmunoprecipitation (co-IP) experiments of PCDH1 and SMAD3 in HEK293T and 16HBE14o(-) (16HBE) cell lines. Activity of a SMAD3-driven luciferase reporter gene in response to TGF-?1 was measured in BEAS-2B cells transfected with PCDH1 and in 16HBE cells transfected with PCDH1-small-interfering RNA (siRNA). TGF-?1-induced gene expression was quantified in BEAS-2B clones overexpressing PCDH1 and in human primary bronchial epithelial cells (PBECs) transfected with PCDH1-siRNA. We confirm PCDH1 and SMAD3 interactions by Y2H and by co-IP in HEK293T cells overexpressing both proteins, and at endogenous protein levels in 16HBE cells. TGF-?-induced activation of a SMAD3-driven reporter was reduced by exogenous PCDH1 in BEAS2B cells, whereas it was increased by siRNA-mediated knockdown of endogenous PCDH1 in 16HBE cells. Overexpression of PCDH1 suppressed expression of TGF-? target genes in BEAS-2B cells, whereas knockdown of PCDH1 in human PBECs increased TGF-?-induced gene expression. In conclusion, we demonstrate that PCDH1 binds to SMAD3 and regulates its activation by TGF-? signaling in bronchial epithelial cells. We propose that PCDH1 and SMAD3 act in a single pathway in asthma susceptibility that affects sensitivity of the airway epithelium to TGF-?.

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More information

Accepted/In Press date: 22 July 2015
Published date: 1 October 2015
Keywords: Mothers against decapentaplegic homolog-3, airway epithelium, functional genetics, asthma susceptibility, transforming growth factor-?-induced gene expression
Organisations: Human Development & Health, Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 388342
URI: https://eprints.soton.ac.uk/id/eprint/388342
ISSN: 1040-0605
PURE UUID: 916c43d7-530a-41c1-8096-70b69d196fac
ORCID for John Holloway: ORCID iD orcid.org/0000-0001-9998-0464

Catalogue record

Date deposited: 24 Feb 2016 11:38
Last modified: 17 Jul 2019 01:08

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