AT-hook DNA-binding motif-containing protein one knockdown downregulates EWS-FLI1 transcriptional activity in Ewing's sarcoma cells

Takao Kitagawa; Daiki Kobayashi; Byron Baron; Hajime Okita; Tatsuo Miyamoto; Rie Takai; Durga Paudel; Tohru Ohta; Yoichi Asaoka; Masayuki Tokunaga; Koji Nakagawa; Makoto Furutani-Seiki; Norie Araki; Yasuhiro Kuramitsu; Masanobu Kobayashi

doi:10.1371/journal.pone.0269077

AT-hook DNA-binding motif-containing protein one knockdown downregulates EWS-FLI1 transcriptional activity in Ewing's sarcoma cells

PLoS One. 2022 Oct 4;17(10):e0269077. doi: 10.1371/journal.pone.0269077. eCollection 2022.

Authors

Affiliations

¹ Advanced Research Promotion Center, Health Sciences University of Hokkaido, Kanazawa, Ishikari-Tobetsu, Hokkaido, Japan.
² Department of Omics and Systems Biology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan.
³ Department of Tumor Genetics and Biology, Faculty of Life Sciences, Kumamoto University, Kumamoto-Shi, Kumamoto, Japan.
⁴ Center for Molecular Medicine and Biobanking, University of Malta, Msida, Malta.
⁵ Division of Diagnostic Pathology, Keio University School of Medicine, Shinano, Shinjuku-ku, Tokyo, Japan.
⁶ Department of Molecular and Cellular Physiology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan.
⁷ Department of Systems Biochemistry in Pathology and Regeneration, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan.
⁸ Department of Obstetrics and Gynecology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan.

Abstract

Ewing's sarcoma is the second most common bone malignancy in children or young adults and is caused by an oncogenic transcription factor by a chromosomal translocation between the EWSR1 gene and the ETS transcription factor family. However, the transcriptional mechanism of EWS-ETS fusion proteins is still unclear. To identify the transcriptional complexes of EWS-ETS fusion transcription factors, we applied a proximal labeling system called BioID in Ewing's sarcoma cells. We identified AHDC1 as a proximal protein of EWS-ETS fusion proteins. AHDC1 knockdown showed a reduced cell growth and transcriptional activity of EWS-FLI1. AHDC1 knockdown also reduced BRD4 and BRG1 protein levels, both known as interacting proteins of EWS-FLI1. Our results suggest that AHDC1 supports cell growth through EWS-FLI1.

MeSH terms

Cell Cycle Proteins / metabolism
Child
DNA
DNA-Binding Proteins / genetics
Gene Expression Regulation, Neoplastic
Humans
Nuclear Proteins / genetics
Oncogene Proteins, Fusion / metabolism
Proto-Oncogene Protein c-fli-1 / genetics
Proto-Oncogene Protein c-fli-1 / metabolism
RNA-Binding Protein EWS / genetics
RNA-Binding Protein EWS / metabolism
Sarcoma, Ewing* / pathology
Transcription Factors / genetics
Transcription Factors / metabolism

Substances

AHDC1 protein, human
BRD4 protein, human
Cell Cycle Proteins
DNA-Binding Proteins
EWS-FLI fusion protein
Nuclear Proteins
Oncogene Proteins, Fusion
Proto-Oncogene Protein c-fli-1
RNA-Binding Protein EWS
Transcription Factors
DNA

Grants and funding

This research was supported by Grant-in-Aid for Scientific Research (C) (Grant number: 18K07199) from the Japan Society for the Promotion of Science (to TK) and the Specific Research Grants (Grant number: 2019081196) from the Takeda Science Foundation (to MFS).