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Arabidopsis Blue Light Receptor Phototropin 1 Undergoes Blue Light-Induced Activation in membrane microdomains

Molecular Plant 2018年第6期11卷 846-859页

作者： Yiqun Xue Jingjing Xing Yinglang Wan Xueqin Lv Lusheng Fan Yongdeng Zhang Kai Song Li Wang Xiaohua Wang Xin Deng Frantisek Baluska John M. Christie Jinxing Lin Institute of Botany Chinese Academy of Sciences Beijing 100093 China University of Chinese Academy of Sciences Beijing 100049 China Department of Botany and Plant Sciences University of California Riverside CA 92521 USA Department of Cell Biology Yale University School of Medicine New Haven CT 06520 USA Institute of Cellular and Molecular Botany University of Bonn 53115 Bonn Germany Institute of Molecular Cell and Systems Biology University of Glasgow Bower Building Glasgow G12 8QQ UK College of Biological Sciences and Biotechnology Beijing Forestry University Beijing 100083 China

Phototropin （phot）-mediated signaling initiated by blue light （BL） plays a critical role in optimizing photosyn- thetic light capture at the plasma membrane （PM） in plants. However, the mechanisms underlying the regu- lation of phot activity at the PM in response to BL remain largely unclear. In this study, by single-particle tracking and stepwise photobleaching analysis of photl-GFP proteins we demonstrated that in the dark photl proteins remain in an inactive state and mostly exist as monomers. Dimerization and the diffusion rate of photl-GFP increased in a dose-dependent manner in response to BL. In contrast, BL did not affect the lateral diffusion of kinase-inactive photlD806N-GFP but did enhance its dimerization, suggesting that photl dimerization is independent of phosphorylation. Forster resonance energy transfer-fluorescence life- time imaging microscopy analysis revealed that the interaction between photl-GFP and a marker of sterol- rich lipid environments, AtRem1.3-mCherry, was enhanced with increased time of BL treatment. However, this BL-dependent interaction was not obvious in plants co-expressing phot1D806N-GFP and AtRem1.3- mCherry, indicating that BL facilitates the translocation of functional photl-GFP into AtRem1.3-1abeled microdomains to activate phot-mediated signaling. Conversely, sterol depletion attenuated photl-GFP dynamics, dimerization, and phosphorylation. Taken together, these results indicate that membrane micro- domains act as organizing platforms essential for the proper function of activated photl at the PM.

关键词： photl VA-TIRFM spatiotemporal dynamics blue light signaling membrane microdomains

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Spatiotemporal Dynamics of the BRI1 Receptor and its Regulation by membrane microdomains in Living Arabidopsis Cells

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Molecular Plant 2015年第9期8卷 1334-1349页

作者： Li Wang Hong Li Xueqin Lv Tong Chen Ruili Li Yiqun xuei Jianjun Jiang Biao Jin Frantisek Baluska Jozef Samaj Xuelu Wang Jinxing Lin Key Laboratory of Ptant Resources Institute of Botany Chinese Academy of Sciences Beijing 100093 China College of Horticulture and Plant Protection Yangzhou University Yangzhou 225009 China College of Biological Sciences and Biotechnology Beijing Forestry University Beijing 100083 China State Key Laboratory of Genetic Engineering and Institute of Plant Biology School of Life Sciences Fudan University Shanghai 200433 China Institute of Cellular and Molecular Botany University of Bonn Kirschallee 1 D-53115 Bonn Germany Department of Cell Biology Palacky University OIomouc Olomouc 78371 Czech Republic College of Life Science and Technology Huazhong Agricultural University Wuhan 430070 China These authors contributed equally to the article.

The major brassinosteroid （BR） receptor of Arabidopsis BRASSINOSTEROID INSENSITIVE1 （BRI1） plays fundamental roles in BR signaling, but the molecular mechanisms underlying the effects of BR on BRI1 internalization and assembly state remain unclear. Here, we applied variable angle total internal reflection fluorescence microscopy and fluorescence cross-correlation spectroscopy to analyze the dynamics of GFP-tagged BRII. We found that, in response to BR, the degree of co-localization of BRI1-GFP with AtFIotl-mCherry increased, and especially BR stimulated the membrane microdomain-associated pathway of BRI1 internalization. We also verified these observations in endocytosis-defective chc2-1 mutants and the AtFIotl amiRNA 15-5 lines. Furthermore, examination of the phosphorylation status of bril-EMS-suppressor 1 and measurement of BR-responsive gene expression revealed that membrane microdomains affect BR signaling. These results suggest that BR promotes the partitioning of BRI1 into functional membrane microdomains to activate BR signaling.

关键词： BRI1 BR signaling endocytosis membrane microdomains spatiotemporal dynamics

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