Current Affiliation Organization 【 display / non-display 】

Current Affiliation Organization 【 display / non-display 】

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External Career 【 display / non-display 】

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Research Areas 【 display / non-display 】

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Published Papers 【 display / non-display 】

• Simultaneous measurement of nascent transcriptome and translatome using 4-thiouridine metabolic RNA labeling and translating ribosome affinity purification

  Imai Hirotatsu, Utsumi Daisuke, Torihara Hidetsugu, Takahashi Kenzo, Kuroyanagi Hidehito, Yamashita Akio

  Nucleic Acids Research ( Oxford University Press )  51 ( 14 ) 1 - 13   2023.06

  Type of publication: Research paper (scientific journal)

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  Regulation of gene expression in response to various biological processes, including extracellular stimulation and environmental adaptation requires nascent RNA synthesis and translation. Analysis of the coordinated regulation of dynamic RNA synthesis and translation is required to determine functional protein production. However, reliable methods for the simultaneous measurement of nascent RNA synthesis and translation at the gene level are limited. Here, we developed a novel method for the simultaneous assessment of nascent RNA synthesis and translation by combining 4-thiouridine (4sU) metabolic RNA labeling and translating ribosome affinity purification (TRAP) using a monoclonal antibody against evolutionarily conserved ribosomal P-stalk proteins. The P-stalk-mediated TRAP (P-TRAP) technique recovered endogenous translating ribosomes, allowing easy translatome analysis of various eukaryotes. We validated this method in mammalian cells by demonstrating that acute unfolded protein response (UPR) in the endoplasmic reticulum (ER) induces dynamic reprogramming of nascent RNA synthesis and translation. Our nascent P-TRAP (nP-TRAP) method may serve as a simple and powerful tool for analyzing the coordinated regulation of transcription and translation of individual genes in various eukaryotes.

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• Roles of Pyk2 in signal transduction after gonadotropin-releasing hormone receptor stimulation.

  Okitsu-Sakurayama S, Higa-Nakamine S, Torihara H, Higashiyama S, Yamamoto H

  Journal of cellular physiology ( Journal of Cellular Physiology )  236 ( 4 ) 3033 - 3043   2021.04 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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• Activation of Pyk2 by CaM kinase II in cultured hypothalamic neurons and gonadotroph cells.

  Okitsu-Sakurayama S, Higa-Nakamine S, Torihara H, Takahashi H, Higashiyama S, Yamamoto H

  Journal of cellular physiology ( Journal of Cellular Physiology )  234 ( 5 ) 6865 - 6875   2019.05 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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• Phosphorylation of epidermal growth factor receptor at serine 1047 in cultured lung alveolar epithelial cells by bradykinin B2 receptor stimulation.

  Izumi S, Higa-Nakamine S, Nishi H, Torihara H, Uehara A, Sugahara K, Kakinohana M, Yamamoto H

  Pulmonary pharmacology & therapeutics ( Pulmonary Pharmacology and Therapeutics )  48   53 - 61   2018.02 [ Peer Review Accepted ]

  Type of publication: Research paper (scientific journal)

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Grant-in-Aid for Scientific Research 【 display / non-display 】

Grant-in-Aid for Scientific Research 【 display / non-display 】

• Grant-in-Aid for Scientific Research(C)

  Project Year: 2020.04  -  2023.03 

  Direct: 3,300,000 (YEN)  Overheads: 990,000 (YEN)  Total: 4,290,000 (YEN)

• Grant-in-Aid for Young Scientists(B)

  Project Year: 2018.04  -  2020.03 

  Direct: 3,200,000 (YEN)  Overheads: 960,000 (YEN)  Total: 4,160,000 (YEN)

• Role of ErbB4 receptor in schizophrenia

  Grant-in-Aid for Scientific Research(C)

  Project Year: 2017.04  -  2020.03 

  Investigator(s): Nakamine Sayomi 

  Direct: 3,900,000 (YEN)  Overheads: 5,070,000 (YEN)  Total: 1,170,000 (YEN)

  　View Summary

  The receptor for gonadotropin-releasing hormone (GnRH) belongs to the G-protein coupled receptors, and is highly expressed in hypothalamic neurons, as well as in anterior pituitary gonadotrophs. In our previous study, we found that GT1-7 cells expressed ErbB4 as well as EGFR, and GnRH treatment induced the cleavage of ErbB4 to suppress the function of ErbB4. In the present study, we found that ErbB4 was expressed in undifferentiated gonadotroph aT3-1 cells and GnRH induced the cleavage of ErbB4. We found that stimulation of the GnRH receptor activated protein kinase D1 (PKD1), and PKD1 was involved in the Fyn-mediated activation of proline-rich tyrosine kinase 2 (Pyk2) in GT1-7 cells. Pyk2 indicated that tyrosine 402 (Tyr402) was phosphorylated both by autophosphorylation and by Fyn, whereas Tyr579 was phosphorylated mainly by Fyn. Our present data may suggest that fully activated Pyk2 dissociates from Fyn after Fyn-mediated phosphorylation of Pyk2 at unknown sites.

• Activation of tyrosine kinases by CaM kinase family in neurons

  Grant-in-Aid for Scientific Research(C)

  Project Year: 2016.04  -  2019.03 

  Investigator(s): Yamamoto Hideyuki 

  Direct: 3,800,000 (YEN)  Overheads: 4,940,000 (YEN)  Total: 1,140,000 (YEN)

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  The receptor of gonadotropin-releasing hormone (GnRH) belongs to the G-protein-coupled receptors (GPCR), and the stimulation of the receptor activates proline-rich tyrosine kinase 2 (Pyk2) and EGFR family. In the present study, we examined the signal transduction pathways after GnRH receptor stimulation using cultured hypothalamic neurons (GT1-7 cells). We first found that GnRH activated CaM kinase II and protein kinase D (PKD), both of which belong to CaM kinase family. And then, we examined the possibility that these two protein kinases were involved in the activation of Pyk2 and EGFR family. Overexpression, knockdown, and inhibitor studies revealed that CaM kinase II and PKD activated Pyk2 through enhancement of interaction of Pyk2 and Fyn. And then, activated Pyk2 induced activation of EGFR. The present study will contribute to the understanding of the physiological roles of CaM kinase family in neurons.