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Elucidation of the role of LMO2 in human erythroid cells
Open accessAbstract
LIM-only protein 2 (LMO2) is a non-DNA-binding component of a protein complex containing master regulators of hematopoiesis, including GATA-1, SCL/TAL1, and LDB1. However, the role of LMO2 in human erythroid differentiation is unclear. LMO2 knockdown in hemin-treated K562 cells reduced the benzidine-positive cell ratio, suggesting that LMO2 retards hemin-mediated K562 cell differentiation. Microarray analysis using K562 cells after siRNA-mediated LMO2 knockdown indicated that 177 and 78 genes were upregulated and downregulated (>1.5-fold), respectively. The downregulated gene ensemble contained prototypical erythroid genes (HBB, SLC4A1). Whereas LMO2 knockdown did not affect GATA-1 or SCL/TAL1 expression, it resulted in significantly reduced chromatin occupancy of GATA-1, SCL/TAL1, and LDB1 at the β-globin locus control region and SLC4A1 locus in both K562 cells and human induced pluripotent stem cell-derived erythroid cells. Introduction of GATA-1 mutations, shown to impair direct interaction with LMO2, significantly diminished chromatin occupancy. On the other hand, knockdown of either SCL/TAL1 or LDB1 also resulted in significantly reduced chromatin occupancy of GATA-1 at endogenous loci, suggesting that impaired assembly of these components also affects GATA-1 chromatin occupancy. In an ex vivo model of erythroid differentiation from CD34(+) cells, LMO2 protein level peaked on day 5 and decreased at later stages of differentiation. The LMO2 expression pattern was similar to those of GATA-1 and SCL/TAL1. Furthermore, shRNA-mediated LMO2 knockdown in primary erythroblasts suggested that LMO2 regulates HBB, HBA, and SLC4A1 expression. LMO2 contributes to GATA-1 target gene expression by affecting assembly of the GATA-SCL/TAL1 complex components at endogenous loci.
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“However, simulation results using Paul et al. data16 showed a different phenotype in erythrocyte cells, suggesting that Lmo2 is also crucial for promoting erythroid differentiation (Extended Data Fig. 6d).”
From Dissecting cell identity via network inference and in silico gene perturbation (2023) · cites this paper as [75]
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Metadata
- Authors
- Inoue, A., Fujiwara, T., Okitsu, Y., Katsuoka, Y., Fukuhara, N., Onishi, Y., Ishizawa, K., & Harigae, H.
- Affiliations
- Tohoku University (JP)
- Corresponding author
- Fujiwara, Tohru
- Venue
- Experimental Hematology
- Publisher
- Elsevier BV
- Published
- 2013-12
- Volume
- 41
- Issue
- 12
- Pages
- 1062-1076.e1
- ISSN
- 0301-472X (ISSN-L)
- OpenAlex
- W1983456610
- Cited by
- 15
- References
- 39
- Citations by year
- 2026: 3 · 2024: 1 · 2023: 2 · 2022: 1 · 2021: 3 · 2020: 1 · 2019: 1 · 2017: 3
- FWCI
- 0.00
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Experimental Hematology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Japan Society for the Promotion of Science (23791067)
- RIKEN
- Topics
- Health Sciences › Medicine › Genetics (clinical) › Hemoglobinopathies and Related Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
- Keywords
- LMO2, erythroid differentiation, GATA-1, SCL/TAL1, LDB1, chromatin occupancy, beta-globin locus control region, band 3, hemin, K562 cells, erythroblasts, gene expression regulation
- MeSH terms
- Cell Differentiation, Erythroblasts, Flow Cytometry, Gene Expression Regulation, Hemin, Humans, Proto-Oncogene Proteins, Blotting, Western, Gene Expression Profiling, Erythroid Cells, Cell Line, Tumor, Adaptor Proteins, Signal Transducing, Gene Knockdown Techniques, LIM Domain Proteins, Real-Time Polymerase Chain Reaction
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