Direct repression of anoctamin 1 (ANO1) gene transcription by Gli proteins

• Published in: The FASEB journal Vol. 33; no. 5; pp. 6632 - 6642
• Main Authors: Mazzone, Amelia, Gibbons, Simon J., Eisenman, Seth T., Strege, Peter R., Zheng, Tenghao, D'Amato, Mauro, Ordog, Tamas, Fernandez‐Zapico, Martin E., Farrugia, And Gianrico
• Format: Journal Article
• Language: English
• Published: United States Federation of American Societies for Experimental Biology 01-05-2019
• Subjects: Anoctamin-1 - biosynthesis; Anoctamin-1 - genetics; Calcium - metabolism; Calcium Signaling; calcium‐activated chloride currents; gastrointestinal tract; Gene Expression Regulation; HEK293 Cells; Humans; ion channels; Irritable Bowel Syndrome - genetics; Irritable Bowel Syndrome - metabolism; Medicin och hälsovetenskap; Mutation; Neoplasm Proteins - biosynthesis; Neoplasm Proteins - genetics; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Polymorphism, Single Nucleotide; SNP; TMEM16A; Transcription Initiation Site; Transcription, Genetic; Zinc Finger Protein GLI1 - genetics; Zinc Finger Protein GLI1 - metabolism; Zinc Finger Protein Gli2 - genetics; Zinc Finger Protein Gli2 - metabolism; calcium-activated chloride currents; gastrointestinal tract; TMEM16A; SNP; ion channels
• ISSN: 0892-6638; 1530-6860; 1530-6860
• DOI: 10.1096/fj.201802373R

ABSTRACTThe 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