The effect of PU.1 knockdown on gene expression and function of mast cells

Oda, Yoshihito; Kasakura, Kazumi; Fujigaki, Izumi; Kageyama, Azusa; Okumura, Ko; Ogawa, Hideoki; Yashiro, Takashi; Nishiyama, Chiharu

doi:10.1038/s41598-018-19378-y

Cited by 17 publications

(13 citation statements)

References 29 publications

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“…The involvement of the GATA factors in the regulation of the human MS4A2 and mouse Ms4a2 genes has been reported in previous studies as well (20)(21)(22). Interestingly, a recent study showed that Ms4a2 gene expression is also affected by small interfering RNA (siRNA)-mediated knockdown (KD) of PU.1 in mouse BMMCs (23). Thus, it is speculated that the mouse Ms4a2 gene might be regulated cooperatively by the GATA factors and PU.1 in mast cells.…”

mentioning

confidence: 73%

GATA2 and PU.1 Collaborate To Activate the Expression of the Mouse Ms4a2 Gene, Encoding FcεRIβ, through Distinct Mechanisms

Ohmori

Ishijima

Numata

et al. 2019

Molecular and Cellular Biology

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GATA factors GATA1 and GATA2 and ETS factor PU.1 are known to function antagonistically during hematopoietic development. In mouse mast cells, however, these factors are coexpressed and activate the expression of the Ms4a2 gene encoding the β chain of the high-affinity IgE receptor (FcεRI). The present study showed that these factors cooperatively regulate Ms4a2 gene expression through distinct mechanisms. Although GATA2 and PU.1 contributed almost equally to Ms4a2 gene expression, gene ablation experiments revealed that simultaneous knockdown of both factors showed neither a synergistic nor an additive effect. A chromatin immunoprecipitation analysis showed that they shared DNA binding to the +10.4-kbp region downstream of the Ms4a2 gene with chromatin looping factor LDB1, whereas the proximal −60-bp region was exclusively bound by GATA2 in a mast cell-specific manner. Ablation of PU.1 significantly reduced the level of GATA2 binding to both the +10.4-kbp and −60-bp regions. Surprisingly, the deletion of the +10.4-kbp region by genome editing completely abolished the Ms4a2 gene expression as well as the cell surface expression of FcεRI. These results suggest that PU.1 and LDB1 play central roles in the formation of active chromatin structure whereas GATA2 directly activates the Ms4a2 promoter.

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mentioning

confidence: 73%

GATA2 and PU.1 Collaborate To Activate the Expression of the Mouse Ms4a2 Gene, Encoding FcεRIβ, through Distinct Mechanisms

Ohmori

Ishijima

Numata

et al. 2019

Molecular and Cellular Biology

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“…The EICE element was changed from 59-TTTCACTTCC-39 to 59-TTGCTAGCCC-39 by a PrimeSTAR Mutagenesis Basal Kit (Takara Bio) to generate a mutant reporter plasmid. Transfection of HEK293T cells and determination of luciferase activity and b-galactosidase activity were performed as described in our previous reports (13,16).…”

Section: Luciferase Assaymentioning

confidence: 99%

“…Probe DNA and transcription factor proteins were prepared as previously described (12,16). Fluorescence of electrophoresis gels was detected using an image analyzer, Typhoon FLA7000 (GE Healthcare).…”

Section: Emsamentioning

confidence: 99%

The Transcription Factors PU.1 and IRF4 Determine Dendritic Cell–Specific Expression of RALDH2

Yashiro

Yamaguchi

Watanuki

et al. 2018

The Journal of Immunology

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RALDH2 expressed in dendritic cells (DCs) plays a critical role in the development of regulatory T cells in mesenteric lymph nodes. Despite the importance of RALDH2 in intestinal immunity, little is known about the mechanism of DC-specific expression of RALDH2. In the current study, we focused on the hematopoietic cell–specific transcription factors PU.1 and IRF4 as the determinants of Aldh1a2 gene expression. The mRNA level of Aldh1a2, and subsequently the enzyme activity, were decreased by knockdown of PU.1 and IRF4 in bone marrow–derived DCs (BMDCs) of BALB/c mice. Chromatin immunoprecipitation assays showed that PU.1 and IRF4 bound to the Aldh1a2 gene ∼2 kb upstream from the transcription start site in BMDCs. A reporter assay and an EMSA revealed that the Aldh1a2 promoter was synergistically transactivated by a heterodimer composed with PU.1 and IRF4 via the EICE motif at −1961/−1952 of the gene. The effect of small interfering RNAs for Spi1 and Irf4 and specific binding of PU.1 and IRF4 on the Aldh1a2 gene were also observed in DCs freshly isolated from spleen and mesenteric lymph nodes, respectively. GM-CSF stimulation upregulated the Aldh1a2 transcription in Flt3 ligand–generated BMDCs, in which the IRF4 expression and the PU.1 recruitment to the Aldh1a2 promoter were enhanced. We conclude that PU.1 and IRF4 are transactivators of the Aldh1a2 gene in vitro and ex vivo.

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“…PU.1 is indispensable for mast cell homeostasis and differentiation evident by failure of PU.1 knock out fetal liver cells to differentiate into mast cells in the presence of SCF and IL3 (73). In a study involving PU.1 siRNA knock down in BMMCs, a significant reduction in the IgE mediated mast cell activation was accompanied with suppression of Syk and FcεRIβ mRNA levels (40). Studies involving siRNA mediated downregulation PU.1, GATA1, and GATA2 in the human mast cell line LAD2 showed a significant reduction in the expression of FcεRI which was further supported by suppressed degranulation from LAD2 cells (74).…”

Section: Ets Family: Ehf and Pu1mentioning

confidence: 99%

Transcription Factors in the Development and Pro-Allergic Function of Mast Cells

Srivastava

Kaplan

2021

Front.Allergy

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Mast cells (MCs) are innate immune cells of hematopoietic origin localized in the mucosal tissues of the body and are broadly implicated in the pathogenesis of allergic inflammation. Transcription factors have a pivotal role in the development and differentiation of mast cells in response to various microenvironmental signals encountered in the resident tissues. Understanding the regulation of mast cells by transcription factors is therefore vital for mechanistic insights into allergic diseases. In this review we summarize advances in defining the transcription factors that impact the development of mast cells throughout the body and in specific tissues, and factors that are involved in responding to the extracellular milieu. We will further describe the complex networks of transcription factors that impact mast cell physiology and expansion during allergic inflammation and functions from degranulation to cytokine secretion. As our understanding of the heterogeneity of mast cells becomes more detailed, the contribution of specific transcription factors in mast cell-dependent functions will potentially offer new pathways for therapeutic targeting.

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The effect of PU.1 knockdown on gene expression and function of mast cells

Cited by 17 publications

References 29 publications

GATA2 and PU.1 Collaborate To Activate the Expression of the Mouse Ms4a2 Gene, Encoding FcεRIβ, through Distinct Mechanisms

GATA2 and PU.1 Collaborate To Activate the Expression of the Mouse Ms4a2 Gene, Encoding FcεRIβ, through Distinct Mechanisms

The Transcription Factors PU.1 and IRF4 Determine Dendritic Cell–Specific Expression of RALDH2

Transcription Factors in the Development and Pro-Allergic Function of Mast Cells

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