Mino M, Kubota M, Nogi T, Zhang S, Inoue M
Graduate School of Agriculture, Kyoto Prefectural University, 1-5 Hangi-cho, Shimogamo, Sakyo-ku, Kyoto 606-8255, Japan.
Plant Biol (Stuttg). 2007 May;9(3):366-73. doi: 10.1055/s-2006-924725. Epub 2007 Jan 19.
A cultured cell line, GTH4 (Nicotiana gossei Domin x N. tabacum L.), which exhibits hybrid lethality, died at 26 degrees C, but not at 37 degrees C. Pharmacological experiments using inhibitors of protein phosphatases and protein kinases indicated the involvement of a protein kinase signalling pathway in the cell death process. Immunoblot analysis revealed that salicylic acid-induced protein kinase (SIPK) was phosphorylated soon after the shift in temperature from 37 degrees C to 26 degrees C. Cultured cells of the hybrid of N. gossei x transgenic N. tabacum harboring a steroid (dexamethasone; DEX)-inducible NtMEK2 (DD) or NtMEK2 (KR), constitutively active and inactive forms of NtMEK2, respectively, were established. Induction of NtMEK2 (DD) by DEX in the hybrid cells induced the activation of SIPK, the generation of hydrogen peroxide (H (2)O (2)), and cell death at 37 degrees C. The activation of SIPK, generation of H (2)O (2), and cell death at 26 degrees C were compromised by DEX treatment in hybrid cells harbouring NtMEK2 (KR). This study provides evidence for the involvement of MAPK signalling in the regulation of cell death in hybrids.
一种表现出杂种致死性的培养细胞系GTH4(粉蓝烟草(Nicotiana gossei Domin)×烟草(N. tabacum L.)),在26℃时死亡,但在37℃时不会。使用蛋白磷酸酶和蛋白激酶抑制剂的药理学实验表明,蛋白激酶信号通路参与了细胞死亡过程。免疫印迹分析显示,在温度从37℃转变为26℃后不久,水杨酸诱导的蛋白激酶(SIPK)就被磷酸化。构建了粉蓝烟草与携带类固醇(地塞米松;DEX)诱导型NtMEK2(DD)或NtMEK2(KR)(分别为NtMEK2的组成型活性形式和非活性形式)的转基因烟草杂交的培养细胞。在杂种细胞中,DEX诱导NtMEK2(DD)会诱导SIPK的激活、过氧化氢(H₂O₂)的产生以及在37℃时的细胞死亡。在携带NtMEK2(KR)的杂种细胞中进行DEX处理后,SIPK的激活、H₂O₂的产生以及在26℃时的细胞死亡均受到抑制。本研究为丝裂原活化蛋白激酶(MAPK)信号传导参与杂种细胞死亡调控提供了证据