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首页> 外文期刊>Nucleus >Identification of candidates for interacting partners of the tail domain of DcNMCP1, a major component of the Daucus carota nuclear lamina-like structure
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Identification of candidates for interacting partners of the tail domain of DcNMCP1, a major component of the Daucus carota nuclear lamina-like structure

机译：鉴定DcNMCP1尾结构域的相互作用伙伴的候选物，DcNMCP1是Daucus胡萝卜核层状结构的主要组成部分

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NMCP/CRWN (NUCLEAR MATRIX CONSTITUENT PROTEIN/CROWDED NUCLEI) is a major component of a protein fibrous meshwork (lamina-like structure) on the plant inner nuclear membrane. NMCP/CRWN contributes to regulating nuclear shape and nuclear functions. An NMCP/CRWN protein in Daucus carota (DcNMCP1) is localized to the nuclear periphery in interphase cells, and surrounds chromosomes in cells in metaphase and anaphase. The N-terminal region and the C-terminal region of DcNMCP1 are both necessary for localizing DcNMCP1 to the nuclear periphery. Here candidate interacting partners of the amino acid position 975–1053 of DcNMCP1 (T975–1053), which is present in the C-terminal region and contains a conserved sequence that plays a role in localizing DcNMCP1 to the nuclear periphery, are screened for. Arabidopsis thaliana nuclear proteins were subjected to far-Western blotting with GST-fused T975–1053 as a probe, and signals were detected at the positions corresponding to ～70, ～40, and ～18?kDa. These ～70, ～40, and ～18?kDa nuclear proteins were identified by mass spectrometry, and subjected to a yeast 2-hybrid (Y2H) analysis with T975–1053 as bait. In this analysis, the ～40?kDa protein ARP7, which is a nuclear actin-related protein possibly involved in regulating chromatin structures, was confirmed to interact with T975–1053. Independently of the far-Western blotting, a Y2H screen was performed using T975–1053 as bait. Targeted Y2H assays confirmed that 3 proteins identified in the screen, MYB3, SINAT1, and BIM1, interact with T975–1053. These proteins might have roles in NMCP/CRWN protein-mediated biologic processes.
机译：NMCP / CRWN（核基质组成蛋白/拥挤核仁）是植物内核膜上蛋白质纤维网状结构（层状结构）的主要成分。 NMCP / CRWN有助于调节核形状和核功能。胡萝卜（Daucus carota）（DcNMCP1）中的NMCP / CRWN蛋白位于间期细胞的核外围，并在中期和后期围绕细胞中的染色体。 DcNMCP1的N端区域和C端区域都是将DcNMCP1定位到核外围所必需的。在此，筛选了存在于C末端区域并包含保守序列的候选相互作用配偶体，该氨基酸位于DcNMCP1的氨基酸位置975–1053（T975–1053）中，该保守序列在将DcNMCP1定位于核外围中发挥作用。用融合了GST的T975-1053作为探针对拟南芥核蛋白进行了远西印迹，并在〜70，〜40和〜18？kDa的位置检测到信号。通过质谱鉴定了这些〜70，〜40和〜18？kDa核蛋白，并以T975–1053为诱饵进行了酵母2-杂交（Y2H）分析。在此分析中，证实〜40？kDa蛋白ARP7是一种可能与调节染色质结构有关的核肌动蛋白相关蛋白，已与T975-1053相互作用。独立于远西印迹法，使用T975-1053作为诱饵进行Y2H筛查。有针对性的Y2H分析证实了筛选中鉴定出的3种蛋白质，MYB3，SINAT1和BIM1与T975–1053相互作用。这些蛋白质可能在NMCP / CRWN蛋白质介导的生物过程中起作用。

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