Vain Thomas, Crowell Elizabeth Faris, Timpano Hélène, Biot Eric, Desprez Thierry, Mansoori Nasim, Trindade Luisa M, Pagant Silvère, Robert Stéphanie, Höfte Herman, Gonneau Martine, Vernhettes Samantha
Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1318, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, F-78000 Versailles, France (T.V., E.F.C., H.T., E.B., T.D., S.P., S.R., H.H., M.G., S.V.);AgroParisTech, Institut Jean-Pierre Bourgin, F-78000 Versailles, France (T.V., E.F.C., H.T., E.B., T.D., H.H., M.G., S.V.); andWageningen University and Research Plant Breeding, Wageningen University and Research Centre, 6708 PD Wageningen, The Netherlands (N.M., L.M.T.).
Institut National de la Recherche Agronomique, Unité Mixte de Recherche 1318, Institut Jean-Pierre Bourgin, Saclay Plant Sciences, F-78000 Versailles, France (T.V., E.F.C., H.T., E.B., T.D., S.P., S.R., H.H., M.G., S.V.);AgroParisTech, Institut Jean-Pierre Bourgin, F-78000 Versailles, France (T.V., E.F.C., H.T., E.B., T.D., H.H., M.G., S.V.); andWageningen University and Research Plant Breeding, Wageningen University and Research Centre, 6708 PD Wageningen, The Netherlands (N.M., L.M.T.)
Plant Physiol. 2014 Aug;165(4):1521-1532. doi: 10.1104/pp.114.241216. Epub 2014 Jun 19.
Plant growth and organ formation depend on the oriented deposition of load-bearing cellulose microfibrils in the cell wall. Cellulose is synthesized by a large relative molecular weight cellulose synthase complex (CSC), which comprises at least three distinct cellulose synthases. Cellulose synthesis in plants or bacteria also requires the activity of an endo-1,4-β-d-glucanase, the exact function of which in the synthesis process is not known. Here, we show, to our knowledge for the first time, that a leaky mutation in the Arabidopsis (Arabidopsis thaliana) membrane-bound endo-1,4-β-d-glucanase KORRIGAN1 (KOR1) not only caused reduced CSC movement in the plasma membrane but also a reduced cellulose synthesis inhibitor-induced accumulation of CSCs in intracellular compartments. This suggests a role for KOR1 both in the synthesis of cellulose microfibrils and in the intracellular trafficking of CSCs. Next, we used a multidisciplinary approach, including live cell imaging, gel filtration chromatography analysis, split ubiquitin assays in yeast (Saccharomyces cerevisiae NMY51), and bimolecular fluorescence complementation, to show that, in contrast to previous observations, KOR1 is an integral part of the primary cell wall CSC in the plasma membrane.
植物的生长和器官形成依赖于细胞壁中承载负荷的纤维素微纤丝的定向沉积。纤维素由一个相对分子质量较大的纤维素合酶复合体(CSC)合成,该复合体至少包含三种不同的纤维素合酶。植物或细菌中的纤维素合成还需要一种内切-1,4-β-D-葡聚糖酶的活性,其在合成过程中的具体功能尚不清楚。在此,据我们所知,我们首次表明,拟南芥中膜结合的内切-1,4-β-D-葡聚糖酶KORRIGAN1(KOR1)的一个渗漏突变不仅导致CSC在质膜中的移动减少,还导致纤维素合成抑制剂诱导的CSC在细胞内区室中的积累减少。这表明KOR1在纤维素微纤丝的合成以及CSC的细胞内运输中均发挥作用。接下来,我们采用了多学科方法,包括活细胞成像、凝胶过滤色谱分析、酵母(酿酒酵母NMY51)中的分裂泛素分析以及双分子荧光互补,以表明,与之前的观察结果相反,KOR1是质膜中初生细胞壁CSC的一个组成部分。