TY - JOUR
T1 - Direct repression of anoctamin 1 (ANO1) gene transcription by Gli proteins
AU - Mazzone, Amelia
AU - Gibbons, Simon J.
AU - Eisenman, Seth T.
AU - Strege, Peter R.
AU - Zheng, Tenghao
AU - D'Amato, Mauro
AU - Ordog, Tamas
AU - Fernandez-Zapico, Martin E.
AU - Farrugia, Gianrico
N1 - Funding Information:
The authors thank Kristy Zodrow (Enteric NeuroSciences, Mayo Clinic) for assistance in manuscript preparation. This work is supported by U.S. National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases Grants DK57061 (to G.F. and S.J.G.) and DK84567 (to G.F., S.J.G.), the Swedish Research Council (VR) Project 2017-02403 (to M.D.A.), the Health Department of the Basque Government (Grant 2015111133 to M.D.A.), and the Spanish Ministry of Economy and Competitiveness [Instituto de Salud Carlos III (ISCIII) Grant Fondo de Investigación en Salud (FIS) PI17/00308 to M.D.A]. The authors declare no conflicts of interest.
Publisher Copyright:
© FASEB
PY - 2019/5/1
Y1 - 2019/5/1
N2 - The Ca2+-activated Cl− channel, anoctamin 1 (Ano1, also known as transmembrane protein 16A) contributes to intestinal pacemaking, fluid secretion, cellular excitability, and tissue development. The human ANO1 promoter contains binding sites for the glioma-associated oncogene (Gli) proteins. We investigated regulation of ANO1 transcription by Gli. ANO1 promoter activity was determined using a luciferase reporter system. Binding and functional effects of Glis on ANO1 transcription and expression were demonstrated by chromatin immunoprecipitation, small interfering RNA knockdown, PCR, immunolabeling, and recordings of Ca2+-activated Cl− currents in human embryonic kidney 293 (HEK293) cells. Results from previous genome-wide association studies were used to test ANO1 promoter polymorphisms for association with disease. Gli1 and Gli2 bound to the promoter and repressed ANO1 transcription. Repression depended on Gli binding to a site close to the ANO1 transcriptional start site. Mutation of this site prevented Gli binding and transcriptional repression. Knockdown of Gli expression and inhibition of Gli activity increased expression of ANO1 RNA and Ca2+-activated Cl− currents in HEK293 cells. A single-nucleotide polymorphism prevented Gli binding and showed association with irritable bowel syndrome. We conclude that Gli1 and Gli2 repress ANO1 by a novel mechanism that is independent of Gli cleavage and that has a role in gastrointestinal function.—Mazzone, A., Gibbons, S. J., Eisenman, S. T., Strege, P. R., Zheng, T., D'Amato, M., Ordog, T., Fernandez-Zapico, M. E., Farrugia, G. Direct repression of anoctamin 1 (ANO1) gene transcription by Gli proteins. FASEB J. 33, 6632–6642 (2019). www.fasebj.org.
AB - The Ca2+-activated Cl− channel, anoctamin 1 (Ano1, also known as transmembrane protein 16A) contributes to intestinal pacemaking, fluid secretion, cellular excitability, and tissue development. The human ANO1 promoter contains binding sites for the glioma-associated oncogene (Gli) proteins. We investigated regulation of ANO1 transcription by Gli. ANO1 promoter activity was determined using a luciferase reporter system. Binding and functional effects of Glis on ANO1 transcription and expression were demonstrated by chromatin immunoprecipitation, small interfering RNA knockdown, PCR, immunolabeling, and recordings of Ca2+-activated Cl− currents in human embryonic kidney 293 (HEK293) cells. Results from previous genome-wide association studies were used to test ANO1 promoter polymorphisms for association with disease. Gli1 and Gli2 bound to the promoter and repressed ANO1 transcription. Repression depended on Gli binding to a site close to the ANO1 transcriptional start site. Mutation of this site prevented Gli binding and transcriptional repression. Knockdown of Gli expression and inhibition of Gli activity increased expression of ANO1 RNA and Ca2+-activated Cl− currents in HEK293 cells. A single-nucleotide polymorphism prevented Gli binding and showed association with irritable bowel syndrome. We conclude that Gli1 and Gli2 repress ANO1 by a novel mechanism that is independent of Gli cleavage and that has a role in gastrointestinal function.—Mazzone, A., Gibbons, S. J., Eisenman, S. T., Strege, P. R., Zheng, T., D'Amato, M., Ordog, T., Fernandez-Zapico, M. E., Farrugia, G. Direct repression of anoctamin 1 (ANO1) gene transcription by Gli proteins. FASEB J. 33, 6632–6642 (2019). www.fasebj.org.
KW - SNP
KW - TMEM16A
KW - calcium-activated chloride currents
KW - gastrointestinal tract
KW - ion channels
UR - http://www.scopus.com/inward/record.url?scp=85065508478&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85065508478&partnerID=8YFLogxK
U2 - 10.1096/fj.201802373R
DO - 10.1096/fj.201802373R
M3 - Article
C2 - 30802137
AN - SCOPUS:85065508478
SN - 0892-6638
VL - 33
SP - 6632
EP - 6642
JO - FASEB Journal
JF - FASEB Journal
IS - 5
ER -