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2,4-D応答にかかわるタンパク質、Small Acidic Protein 1(SMAP1)はCOP9シグナロソーム(CSN)と結合する

SMAP1 that functions in 2,4-D response interacts with COP9 Signalosome

中曽根 光; 川合 真紀*; 鳴海 一成; 内宮 博文*; 大野 豊

Nakasone, Akari; Kawai, Maki*; Narumi, Issei; Uchimiya, Hirofumi*; Ono, Yutaka

われわれが合成オーキシン2,4-Dの応答機構にかかわる因子として同定したSMAP1の機能を解明するために、SMAP1欠失変異体${it aar1}$に完全長及びC末のF/D保存領域を欠失させた${it SMAP1}$${it GFP}$の融合遺伝子(それぞれ${it SMAP1-GFP}$, ${it SMAP1$Delta$F/D-GFP}$)を導入した。その結果、${it SMAP1-GFP}$は根における${it aar1}$の2,4-D感受性を野生型のレベルにまで回復させたが、${it SMAP1$Delta$F/D-GFP}$では回復がみられず、F/D領域がSMAP1の活性に重要であることがわかった。次に、抗GFP-マイクロビーズを用い、SMAP1-GFPと結合するがSMAP1$$Delta$$F/D-GFPとは結合しないタンパク質を精製し、質量分析で調べたところ、そのほとんどがCOP9 Signalosome(CSN)のサブユニットであった。以上の結果から、SMAP1はF/D領域を介してCSNと相互作用し、2,4-D応答を制御している可能性が示唆された。

The ${it Small Acidic Protein 1}$ (${it SMAP1}$) gene is recently identified as the factor that mediates the synthetic auxin 2,4-D response in ${it Arabidopsis}$ root. SMAP1 has the conserved phenylalanine and asparagic acid (F/D) domain, which is highly conserved in both plants and animals. Introduction of ${it SMAP1-GFP}$ to ${it aar1}$ recovered 2,4-D sensitivity of ${it aar1}$ while that of ${it SMAP1$Delta$F/D-GFP}$ did not. Using anti-GFP-MicroBeads, the proteins that form complexes with SMAP1-GFP but not with SMAP1$$Delta$$F/D-GFP were pulled down from the ${it Arabidopsis}$extract and identified by MS analysis. The identified peptide sequences indicated that SMAP1-GFP binds to COP9 Signalosome (CSN). The results implied that SMAP1 regulates 2,4-D response by interacting to CSN through F/D region.

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