Dataset Information

Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gαs protein

ABSTRACT: The mechanism underlying the crosstalk between Gαq-coupled bombesin receptor subtype-3 (BRS3) and Gαi-coupled E-prostanoid 3 receptor (EP3) remains unknown. Here, we report that BRS3 and EP3 form dimers in the membrane of living HEK-293T cells. Interestingly, we found that BRS3-EP3 dimers switched to couple Gαs protein upon PGE2 stimulation, which provoked the accumulation of cAMP and enhanced the phosphorylation of P38. Quantitative proteomics analysis revealed that the activation of BRS3-EP3 dimers was associated with cell migration. B16 melanoma cell line, which endogenously expresses BRS3 and EP3, was selected to investigate the function of BRS3-EP3 dimers. Results demonstrated that the presence of BRS3 inhibited the migration of B16 melanoma cells upon PGE2 stimulation using scratch-wound assay. Utilizing inhibitors of Gαs and P38, we found that BRS3 interacted with EP3 and switched to couple Gαs protein, causing P38 phosphorylation to inhibit F-actin rearrangement and ultimately suppresses cell migration. Our study reveals the crosstalk between the orphan receptor BRS3 and EP3, and provides a potential novel target for disease treatment.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Early Embryonic Cell

DISEASE(S): Melanoma

SUBMITTER: Zeyuan Wang

LAB HEAD: Hua Xiao

PROVIDER: PXD042449 | Pride | 2026-01-05

REPOSITORIES: Pride

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Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gαs protein.

Wang Zeyuan Z Wu Lehao L Guo Miao M Zhu Jianzheng J Zhao Jiaqi J Wu Yan Y Xiao Hua H Zhang Yan Y

Fundamental research 20240425 6

The mechanism underlying the crosstalk between Gαq-coupled bombesin receptor subtype-3 (BRS3) and Gαi-coupled E-prostanoid 3 receptor (EP3) remains unknown. Here, we report that BRS3 and EP3 form dimers in the membrane of living HEK-293T cells. BRS3-EP3 dimers switched to couple Gαs protein upon PGE2 stimulation, which provoked cAMP accumulation and enhanced P38 phosphorylation. Quantitative proteomics analysis revealed that the activation of BRS3-EP3 dimers was ...[more]

PMID: 41466974

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Project description:Recognition of foreign antigens by B cell receptor (BCR) on mature B cells leads to their clonal expansion, which is critically important for the effective host defence of the organism. However, excessive antigenic responses or reaction of B cells against bodyâs own components frequently lead to diverse immune diseases, such as B-cell lymphoma or autoimmunity, that often affect humans. Identification of genes that restrain uncontrolled proliferation of B cells is therefore an important goal towards understanding the origin of such diseases. Here we identify Ptger4 as a negative feedback regulator of B-cell proliferation in response to BCR triggering, and show that its encoded EP4 receptor is a principal molecule conveying the growth-suppressive effect of prostaglandin E2 (PGE2). In controlled in vitro assays, Ptger4-/- B cells showed augmented proliferative response and increased expression of activating genes upon BCR stimulation. Stable knock-down of Ptger4 in B-cell lymphoma markedly accelerated tumour spread in mice, while Ptger4 overexpression yielded significant protection. Lack of Ptger4 rendered mouse B cells completely resistant to proliferation arrest signalled by PGE2, and we find by transcriptional profiling that intrinsic activity of Ptger4 and PGE2-EP4 signalling target a similar set of activating genes. We further show that Ptger4 inhibits mouse B-cell activation in vivo and find it significantly downregulated in human B-cell lymphoma. Our results demonstrate that Ptger4 functions in B cells as a candidate tumour suppressor whose activity is regulated by the presence of PGE2 in the microenvironment. These findings suggest that targeting EP4 receptor for prostaglandin may present a novel strategy for treatment of B-cell diseases. Keywords: time course, cell type comparison, compound treatment design, microarray experiment record B cells were extracted from spleen of Ptger4+/+ and Ptger4-/- mice, and incubated in vitro for the indicated period with or without anti-IgM (Fab')2 antibody fragments and co-treated with or without PGE2. Total RNA was extracted, amplified, labelled with Cy3 or Cy5, and hybridized to mouse 24k oligo-arrays using a dye-swap strategy.

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			Action	DRS
	EHBFCON-1.wiff	Wiff
	EHBFCON-1.wiff.scan	Wiff
	EHBFCON-2.wiff	Wiff
	EHBFCON-2.wiff.scan	Wiff
	EHBFCON-3.wiff	Wiff

Dataset Information

Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gαs protein

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Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gα<sub>s</sub> protein.

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Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gαs protein

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Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gα&lt;sub&gt;s&lt;/sub&gt; protein.

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OmicsDI is part of the ELIXIR infrastructure

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Activation of dimerized BRS3-EP3 suppresses melanoma cell migration through coupling Gα<sub>s</sub> protein.