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

Autoren

Teng Wei Koay
Carina Osterhof
Ilaria MC Orlando
Anna Keppner
Daniel Andre
Schayan Yousefian
María Suárez Alonso
Miguel Correia
Robert Markworth
Johannes Schödel
Thomas Hankeln
David Hoogewijs

DOI

10.1016/j.jbc.2021.100291

ISSN

0021-9258

Zeitschrift

Journal of Biological Chemistry

Sprache

en

Artikelnummer

100291

Paginierung

100291 - 100291

Datum der Veröffentlichung

2021

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1016/j.jbc.2021.100291

Datum der Datenerfassung

2021

Titel

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

Ausgabe der Zeitschrift

296

Datenquelle: Crossref

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.

Addresses

Section of Medicine, Department of Endocrinology, Metabolism and Cardiovascular System, University of Fribourg, Fribourg, Switzerland.

Autoren

Teng Wei Koay
Carina Osterhof
Ilaria MC Orlando
Anna Keppner
Daniel Andre
Schayan Yousefian
María Suárez Alonso
Miguel Correia
Robert Markworth
Johannes Schödel
Thomas Hankeln
David Hoogewijs

DOI

10.1016/j.jbc.2021.100291

eISSN

1083-351X

Externe Identifier

PubMed Identifier: 33453283
PubMed Central ID: PMC7949040

Funding acknowledgements

Deutsche Forschungsgemeinschaft: 387509280
Deutsche Forschungsgemeinschaft: HA 2103/9-1
Deutsche Forschungsgemeinschaft: HO 5837/1-1
Deutsche Forschungsgemeinschaft: SFB 1350
Swiss National Science Foundation: 31003A_173000

Open access

true

ISSN

0021-9258

Zeitschrift

The Journal of biological chemistry

Schlüsselwörter

Lung
Ovary
Testis
Brain
Hela Cells
Cilia
Animals
Cattle
Humans
Globins
Calmodulin-Binding Proteins
Protein Isoforms
Sequence Analysis, RNA
Gene Expression Regulation, Developmental
Binding Sites
Protein Binding
Female
Male
Forkhead Transcription Factors
Enhancer Elements, Genetic
Promoter Regions, Genetic
HEK293 Cells
Molecular Sequence Annotation
Transcriptome
MCF-7 Cells
Gene Ontology
Regulatory Factor X Transcription Factors

Sprache

eng

Medium

Print-Electronic

Online publication date

2021

Open access status

Open Access

Paginierung

100291

Datum der Veröffentlichung

2021

Status

Published

Publisher licence

CC BY

Datum der Datenerfassung

2021

Titel

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

Sub types

Research Support, Non-U.S. Gov't
research-article
Journal Article

Ausgabe der Zeitschrift

296

Files

http://www.jbc.org/article/S0021925821000594/pdf https://europepmc.org/articles/PMC7949040?pdf=render

Datenquelle: Europe PubMed Central

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.

Date of acceptance

2021

Autoren

Teng Wei Koay
Carina Osterhof
Ilaria MC Orlando
Anna Keppner
Daniel Andre
Schayan Yousefian
María Suárez Alonso
Miguel Correia
Robert Markworth
Johannes Schödel
Thomas Hankeln
David Hoogewijs

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/33453283

DOI

10.1016/j.jbc.2021.100291

eISSN

1083-351X

Externe Identifier

PubMed Central ID: PMC7949040

Zeitschrift

J Biol Chem

Schlüsselwörter

CRISPR/cas
bioinformatics
cilia
gene expression
hemoglobin myoglobin
transcription enhancer
transcription factor
transcription regulation
transcriptomics
Animals
Binding Sites
Brain
Calmodulin-Binding Proteins
Cattle
Cilia
Enhancer Elements, Genetic
Female
Forkhead Transcription Factors
Gene Expression Regulation, Developmental
Gene Ontology
Globins
HEK293 Cells
HeLa Cells
Humans
Lung
MCF-7 Cells
Male
Molecular Sequence Annotation
Ovary
Promoter Regions, Genetic
Protein Binding
Protein Isoforms
Regulatory Factor X Transcription Factors
Sequence Analysis, RNA
Testis
Transcriptome

Sprache

eng

Country

United States

Paginierung

100291

PII

S0021-9258(21)00059-4

Datum der Veröffentlichung

2021

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2021

Titel

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

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

296

Datenquelle: PubMed

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

Files

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