Betti Camilla, Vanhoutte Isabelle, Coutuer Silvie, De Rycke Riet, Mishev Kiril, Vuylsteke Marnik, Aesaert Stijn, Rombaut Debbie, Gallardo Rodrigo, De Smet Frederik, Xu Jie, Van Lijsebettens Mieke, Van Breusegem Frank, Inzé Dirk, Rousseau Frederic, Schymkowitz Joost, Russinova Eugenia
Department of Plant Systems Biology, VIB, 9052 Gent, Belgium (C.B., I.V., S.C., R.D.R., K.M., S.A., D.R., M.V.L., F.V.B., D.I., E.R.);Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Gent, Belgium (C.B., I.V., S.C., R.D.R., K.M., S.A., D.R., M.V.L., F.V.B., D.I., E.R.);Switch Laboratory, VIB, 3000 Leuven, Belgium (R.G., F.D.S., J.X., F.R., J.S.);Switch Laboratory, Department of Cellular and Molecular Medicine, University of Leuven, 3000 Leuven, Belgium (R.G., F.D.S., J.X., F.R., J.S); andGnomixx, 9000 Gent, Belgium (M.V.).
Department of Plant Systems Biology, VIB, 9052 Gent, Belgium (C.B., I.V., S.C., R.D.R., K.M., S.A., D.R., M.V.L., F.V.B., D.I., E.R.);Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Gent, Belgium (C.B., I.V., S.C., R.D.R., K.M., S.A., D.R., M.V.L., F.V.B., D.I., E.R.);Switch Laboratory, VIB, 3000 Leuven, Belgium (R.G., F.D.S., J.X., F.R., J.S.);Switch Laboratory, Department of Cellular and Molecular Medicine, University of Leuven, 3000 Leuven, Belgium (R.G., F.D.S., J.X., F.R., J.S); andGnomixx, 9000 Gent, Belgium (M.V.)
Plant Physiol. 2016 Jun;171(2):773-87. doi: 10.1104/pp.16.00335. Epub 2016 May 4.
Protein aggregation is determined by short (5-15 amino acids) aggregation-prone regions (APRs) of the polypeptide sequence that self-associate in a specific manner to form β-structured inclusions. Here, we demonstrate that the sequence specificity of APRs can be exploited to selectively knock down proteins with different localization and function in plants. Synthetic aggregation-prone peptides derived from the APRs of either the negative regulators of the brassinosteroid (BR) signaling, the glycogen synthase kinase 3/Arabidopsis SHAGGY-like kinases (GSK3/ASKs), or the starch-degrading enzyme α-glucan water dikinase were designed. Stable expression of the APRs in Arabidopsis (Arabidopsis thaliana) and maize (Zea mays) induced aggregation of the target proteins, giving rise to plants displaying constitutive BR responses and increased starch content, respectively. Overall, we show that the sequence specificity of APRs can be harnessed to generate aggregation-associated phenotypes in a targeted manner in different subcellular compartments. This study points toward the potential application of induced targeted aggregation as a useful tool to knock down protein functions in plants and, especially, to generate beneficial traits in crops.
蛋白质聚集由多肽序列中短的（5 - 15个氨基酸）易于聚集的区域（APR）决定，这些区域以特定方式自我缔合形成β结构的内含物。在此，我们证明可以利用APR的序列特异性来选择性地敲低植物中具有不同定位和功能的蛋白质。设计了源自油菜素内酯（BR）信号负调节因子、糖原合酶激酶3/拟南芥类SHAGGY激酶（GSK3/ASKs）或淀粉降解酶α-葡聚糖水二激酶的APR的合成易于聚集的肽。APR在拟南芥（Arabidopsis thaliana）和玉米（Zea mays）中的稳定表达诱导了靶蛋白的聚集，分别产生了显示组成型BR反应的植物和淀粉含量增加的植物。总体而言，我们表明可以利用APR的序列特异性在不同亚细胞区室中以靶向方式产生与聚集相关的表型。这项研究指出了诱导靶向聚集作为敲低植物中蛋白质功能，特别是在作物中产生有益性状的有用工具的潜在应用。
