Androglobin gene expression patterns and FOXJ1-dependent regulation indicate its functional association with ciliogenesis

Koay TW, Osterhof C, Orlando IM, Keppner A, Andre D, Yousefian S, Alonso MS, Correia M, Markworth R, Schödel J, Hankeln T, Hoogewijs D (2021)

Publication Type: Journal article

Publication year: 2021

Journal

Journal of Biological Chemistry American Society for Biochemistry and Molecular Biology

Book Volume: 296

Article Number: 100291

DOI: 10.1016/j.jbc.2021.100291

Abstract

Androglobin (ADGB) represents the latest addition to the globin superfamily in metazoans. The chimeric protein comprises a calpain domain and a unique circularly permutated globin domain. ADGB expression levels are most abundant in mammalian testis, but its cell-type-specific expression, regulation, and function have remained unexplored. Analyzing bulk and single-cell mRNA-Seq data from mammalian tissues, we found that—in addition to the testes—ADGB is prominently expressed in the female reproductive tract, lungs, and brain, specifically being associated with cell types forming motile cilia. Correlation analysis suggested coregulation of ADGB with FOXJ1, a crucial transcription factor of ciliogenesis. Investigating the transcriptional regulation of the ADGB gene, we characterized its promoter using epigenomic datasets, exogenous promoter-dependent luciferase assays, and CRISPR/ dCas9-VPR-mediated activation approaches. Reporter gene assays revealed that FOXJ1 indeed substantially enhanced luciferase activity driven by the ADGB promoter. ChIP assays confirmed binding of FOXJ1 to the endogenous ADGB promoter region. We dissected the minimal sequence required for FOXJ1-dependent regulation and fine mapped the FOXJ1 binding site to two evolutionarily conserved regions within the ADGB promoter. FOXJ1 overexpression significantly increased endogenous ADGB mRNA levels in HEK293 and MCF-7 cells. Similar results were observed upon RFX2 overexpression, another key transcription factor in ciliogenesis. The complex transcriptional regulation of the ADGB locus was illustrated by identifying a distal enhancer, responsible for synergistic regulation by RFX2 and FOXJ1. Finally, cell culture studies indicated an ADGB-dependent increase in the number of ciliated cells upon overexpression of the full-length protein, confirming a ciliogenesis-associated role of ADGB in mammals.

Authors with CRIS profile

Johannes Schödel Department of Medicine 4 – Nephrology and Hypertension

Involved external institutions

Johannes Gutenberg-Universität Mainz (JGU) DE Germany (DE) University of Fribourg / Universitè de Fribourg CH Switzerland (CH) Universität Duisburg-Essen (UDE) DE Germany (DE)

How to cite

APA:

Koay, T.W., Osterhof, C., Orlando, I.M., Keppner, A., Andre, D., Yousefian, S.,... Hoogewijs, D. (2021). Androglobin gene expression patterns and FOXJ1-dependent regulation indicate its functional association with ciliogenesis. Journal of Biological Chemistry, 296. https://doi.org/10.1016/j.jbc.2021.100291

MLA:

Koay, Teng Wei, et al. "Androglobin gene expression patterns and FOXJ1-dependent regulation indicate its functional association with ciliogenesis." Journal of Biological Chemistry 296 (2021).

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