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

FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology
Amelia MazzoneGianrico Farrugia

Abstract

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...Continue Reading

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Citations

Jul 11, 2020·Proceedings of the National Academy of Sciences of the United States of America·Alexander KlimovichThomas C G Bosch
Aug 8, 2020·Proceedings of the National Academy of Sciences of the United States of America·Yuuki Obata, Vassilis Pachnis
Aug 9, 2020·The Biochemical Journal·Ezequiel J TolosaMartin E Fernandez-Zapico
Nov 27, 2020·The Biochemical Journal·Alex J Timmis, Natalia A Riobo-Del Galdo

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