Forced FOG1 expression in erythroleukemia cells: Induction of erythroid genes and repression of myelo-lymphoid transcription factor PU.1.

Abstract

The transcription factor GATA-1-interacting protein Friend of GATA-1 (FOG1) is essential for proper transcriptional activation and repression of GATA-1 target genes; yet, the mechanisms by which FOG1 exerts its activating and repressing functions remain unknown. Forced FOG1 expression in human K562 erythroleukemia cells induced the expression of erythroid genes (SLC4A1, globins) but repressed that of GATA-2 and PU.1. A quantitative chromatin immunoprecipitation (ChIP) analysis demonstrated increased GATA-1 chromatin occupancy at both FOG1-activated as well as FOG1-repressed gene loci. However, while TAL1 chromatin occupancy was significantly increased at FOG1-activated gene loci, it was significantly decreased at FOG1-repressed gene loci. When FOG1 was overexpressed in TAL1-knocked down K562 cells, FOG1-mediated activation of HBA, HBG, and SLC4A1 was significantly compromised by TAL1 knockdown, suggesting that FOG1 may require TAL1 to activate GATA-1 target genes. Promoter analysis and quantitative ChIP analysis demonstrated that FOG1-mediated transcriptional repression of PU.1 would be mediated through a GATA-binding element located at its promoter, accompanied by significantly decreased H3 acetylation at lysine 4 and 9 (K4 and K9) as well as H3K4 trimethylation. Our results provide important mechanistic insight into the role of FOG1 in the regulation of GATA-1-regulated genes and suggest that FOG1 has an important role in inducing cells to differentiate toward the erythroid lineage rather than the myelo-lymphoid one by repressing the expression of PU.1.

• Authors: Tohru Fujiwara, Katsuyuki Sasaki, Kei Saito, Shunsuke Hatta, Satoshi Ichikawa, Masahiro Kobayashi, Yoko Okitsu, Noriko Fukuhara, Yasushi Onishi, Hideo Harigae
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 485 Issue 2 Pg. 380-387 (04 01 2017) ISSN: 1090-2104 [Electronic] United States
• PMID: 28216155 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2017 Elsevier Inc. All rights reserved.

Chemical References

• Anion Exchange Protein 1, Erythrocyte
• Basic Helix-Loop-Helix Transcription Factors
• Chromatin
• GATA1 Transcription Factor
• GATA1 protein, human
• Histones
• Nuclear Proteins
• Proto-Oncogene Proteins
• SLC4A1 protein, human
• T-Cell Acute Lymphocytic Leukemia Protein 1
• Trans-Activators
• Transcription Factors
• ZFPM1 protein, human
• proto-oncogene protein Spi-1
• TAL1 protein, human
• Lysine

Topics

• Acetylation
• Anion Exchange Protein 1, Erythrocyte (genetics, metabolism)
• Basic Helix-Loop-Helix Transcription Factors (genetics, metabolism)
• Blotting, Western
• Cell Line
• Chromatin (genetics, metabolism)
• GATA1 Transcription Factor (genetics, metabolism)
• Gene Expression Regulation, Leukemic
• Histones (metabolism)
• Humans
• K562 Cells
• Leukemia, Erythroblastic, Acute (genetics, metabolism, pathology)
• Lysine (metabolism)
• Nuclear Proteins (genetics, metabolism)
• Promoter Regions, Genetic (genetics)
• Protein Binding
• Proto-Oncogene Proteins (genetics, metabolism)
• RNA Interference
• Reverse Transcriptase Polymerase Chain Reaction
• T-Cell Acute Lymphocytic Leukemia Protein 1
• Trans-Activators (genetics, metabolism)
• Transcription Factors (genetics, metabolism)