DBS is activated by EPHB2/SRC signaling-mediated tyrosine phosphorylation in HEK293 cells

Nakano, Susumu; Nishikawa, Masashi; Asaoka, Rina; Ishikawa, Natsuko; Ohwaki, Chisato; Sato, Katsuya; Nagaoka, Hitoshi; Yamakawa, Hisashi; Nagase, Takahiro; Ueda, Hiroshi

doi:10.1007/s11010-019-03552-5

Cited by 3 publications

(2 citation statements)

References 30 publications

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“…Among those, EphB2 was one of the most upregulated and hypomethylated genes and was previously shown to be involved in T cell proliferation 41 , 42 . It was also shown that Dock2 and EphB2 play a role in lymphoid progenitor migration into the thymus 43 , 44 , and that EphB2 signaling activates CDC42-GEF Mcf2l 45 , suggesting EPHB2 signals through the same pathway as DOCK2.…”

Section: Discussionmentioning

confidence: 99%

Extra-hematopoietic immunomodulatory role of the guanine-exchange factor DOCK2

et al. 2022

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Stromal cells interact with immune cells during initiation and resolution of immune responses, though the precise underlying mechanisms remain to be resolved. Lessons learned from stromal cell-based therapies indicate that environmental signals instruct their immunomodulatory action contributing to immune response control. Here, to the best of our knowledge, we show a novel function for the guanine-exchange factor DOCK2 in regulating immunosuppressive function in three human stromal cell models and by siRNA-mediated DOCK2 knockdown. To identify immune function-related stromal cell molecular signatures, we first reprogrammed mesenchymal stem/progenitor cells (MSPCs) into induced pluripotent stem cells (iPSCs) before differentiating these iPSCs in a back-loop into MSPCs. The iPSCs and immature iPS-MSPCs lacked immunosuppressive potential. Successive maturation facilitated immunomodulation, while maintaining clonogenicity, comparable to their parental MSPCs. Sequential transcriptomics and methylomics displayed time-dependent immune-related gene expression trajectories, including DOCK2, eventually resembling parental MSPCs. Severe combined immunodeficiency (SCID) patient-derived fibroblasts harboring bi-allelic DOCK2 mutations showed significantly reduced immunomodulatory capacity compared to non-mutated fibroblasts. Conditional DOCK2 siRNA knockdown in iPS-MSPCs and fibroblasts also immediately reduced immunomodulatory capacity. Conclusively, CRISPR/Cas9-mediated DOCK2 knockout in iPS-MSPCs also resulted in significantly reduced immunomodulation, reduced CDC42 Rho family GTPase activation and blunted filopodia formation. These data identify G protein signaling as key element devising stromal cell immunomodulation.

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Section: Discussionmentioning

confidence: 99%

Extra-hematopoietic immunomodulatory role of the guanine-exchange factor DOCK2

et al. 2022

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“…Recently, we reported that the phosphorylation of Tyr-479 and Tyr-660 on DBS leads to actin cytoskeletal reorganization by EPHB2/SRC signaling. Based on these reports, it is thought that SFKs play an important role in RhoGEF activation (7).…”

Section: Introductionmentioning

confidence: 99%

The Rho guanine nucleotide exchange factor PLEKHG1 is activated by interaction with and phosphorylation by Src family kinase member FYN

Nakano

Nishikawa

Kobayashi

et al. 2022

Journal of Biological Chemistry

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This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

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Loss of tyrosine kinase receptor Ephb2 impairs proliferation and stem cell activity of spermatogonia in culture†

N’Tumba-Byn

Yamada

Seandel

2019

Biology of Reproduction

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Germline stem and progenitor cells can be extracted from the adult mouse testis and maintained long-term in vitro. Yet, the optimal culture conditions for preserving stem cell activity are unknown. Recently, multiple members of the Eph receptor family were detected in murine spermatogonia, but their roles remain obscure. One such gene, Ephb2, is crucial for maintenance of somatic stem cells and was previously found enriched at the level of mRNA in murine spermatogonia. We detected Ephb2 mRNA and protein in primary adult spermatogonial cultures and hypothesized that Ephb2 plays a role in maintenance of stem cells in vitro. We employed CRISPR-Cas9 targeting and generated stable mutant SSC lines with complete loss of Ephb2. The characteristics of Ephb2-KO cells were interrogated using phenotypic and functional assays. Ephb2-KO SSCs exhibited reduced proliferation compared to wild-type cells, while apoptosis was unaffected. Therefore, we examined whether Ephb2 loss correlates with activity of canonical pathways involved in stem cell self-renewal and proliferation. Ephb2-KO cells had reduced ERK MAPK signaling. Using a lentiviral transgene, Ephb2 expression was rescued in Ephb2-KO cells, which partially restored signaling and proliferation. Transplantation analysis revealed that Ephb2-KO SSCs cultures formed significantly fewer colonies than WT, indicating a role for Ephb2 in preserving stem cell activity of cultured cells. Transcriptome analysis of wild-type and Ephb2-KO SSCs identified Dppa4 and Bnc1 as differentially expressed, Ephb2-dependent genes that are potentially involved in stem cell function. These data uncover for the first time a crucial role for Ephb2 signaling in cultured SSCs.

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DBS is activated by EPHB2/SRC signaling-mediated tyrosine phosphorylation in HEK293 cells

Cited by 3 publications

References 30 publications

Extra-hematopoietic immunomodulatory role of the guanine-exchange factor DOCK2

Extra-hematopoietic immunomodulatory role of the guanine-exchange factor DOCK2

The Rho guanine nucleotide exchange factor PLEKHG1 is activated by interaction with and phosphorylation by Src family kinase member FYN

Loss of tyrosine kinase receptor Ephb2 impairs proliferation and stem cell activity of spermatogonia in culture†

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