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CAPPh A Cytoskeleton-Based Localization Assay Reports Protein-Protein Interaction in Living Cells by Fluorescence Microscopy

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Molecular Plant 2017年第11期10卷 1473-1476页

作者： Sulian Lv Huiying Miao Ming Luo Yinxin Li Qiaomei Wang yuh-ru julie lee Bo Liu Department of Plant Biology University of California Davis CA 95616 USA Key Laboratory of Plant Molecular Physiology Institute of Botany Chinese Academy of Sciences Beijing China Department of Horticulture Zhejiang University Hangzhou Zhejiang 310058 China Biotechnology Research Center Southwest University Chongqing 400716 China

Dear EditorProbing protein-protein interaction has become a routine practice in the post genomic era. Multiple in vitro or in vivo techniques have been developed to detect or report direct or indirect interactions of functionally related proteins （Lalonde et al., 2008）. These techniques sometimes are technically challenging, however, because the readout would demand sophisticated detectors and/or complicated calculations. Besides, a common drawback of many of these techniques is they can render inherent false positives to various degrees so that an interaction often cannot be judged unambiguously.

关键词： CAPPh A Cytoskeleton-Based Localization Assay Reports Protein-Protein Interaction Living Cells by Fluorescence Microscopy

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C-terminal tensin-like (CTEN) knockin alleviates cystic kidney defects in Tensin-1 knockout mice

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Genes & Diseases 2023年第3期10卷 643-646页

作者： Chun-Lung Chiu Shiao-Ya Hong Ying Tan yuh-ru julie lee Yi-Ping Shih Clifford GTepper Su Hao Lo Department of Biochemistry and Molecular Medicine University of California-DavisSacramentoCA 95817USA Department of Plant Biology University of California-DavisDavisCA 95616USA

Tensin-1(TNS1)is a 220 kD focal adhesion protein that binds to actin filaments,integrin receptors,small GTPases,tyrosine-phosphorylated proteins,and lipids.1 These binding activities enable TNS1 to link the actin cytoskeleton to integrins and transduce outside-in and inside-out signals at focal adhesion sites,thereby regulating cell attachment,migration,proliferation,and mechanical sensing.1 Lack of TNS1 in kidney epithelial cells results in multiple lumen,instead of single lumen,phenotypes in the three-dimensional(3D)culture.

关键词： kidney Tensin thereby

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Kinesin-4 Functions in Vesicular Transport on Cortical Microtubules and Regulates Cell Wall Mechanics during Cell Elongation in Plants

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Molecular Plant 2015年第7期8卷 1011-1023页

作者： Zhaosheng Kong Motohide Ioki Siobhan Braybrook Shundai Li Zheng-Hua Ye yuh-ru julie lee Takashi Hotta Anny Chang Juan Tian Guangda Wang Bo Liu Department of Plant Biology University of California Davis CA 95616 USA State Key Laboratory of Plant Genomics Institute of Microbiology Chinese Academy of Sciences Beijing 100101 China Sainsbury Laboratory Cambridge University of Cambridge Cambridge CB2 1LR UK Department of Biochemistry and Molecular Biology Pennsylvania State University State College PA 16802 USA Department of Plant Biology University of Georgia Athens GA 30602 USA Present address: Center for Environmental Biology and Ecosystem Studies National Institute for Environmental Studies Onogawa 16-2 Tsukuba Ibaraki305-8506 Japan Present address: Nara Institute of Science and Technology Graduate School of Biological Sciences 8916-5 Takayama Ikoma Nara 630-0192 Japan

In plants, anisotropic cell expansion depends on cortical microtubules that serve as tracks along which macromolecules and vesicles are transported by the motor kinesins of unknown identities. We used cotton （Gossypium hirsutum） fibers that underwent robust elongation to discover kinesins that are involved in cell elongation and found Gh KINESIN-4A expressed abundantly. The motor was detected by immunofluores- cence on vesicle-like structures that were associated with cortical microtubules. In Arabidopsis thaliana, the orthologous motor At KINESIN-4A/FRA1, previously implicated in cellulose deposition during second- ary growth in fiber cells, was examined by live-cell imaging in cells expressing the fluorescently tagged functional protein. The motor decorated vesicle-like particles that exhibit a linear movement along cortical microtubules with an average velocity of 0.89 l^m/min, which was significantly different from those linked to cellulose biosynthesis. We also discovered that At KINESIN-4A/FRA1 and the related At KINESIN-4C play redundant roles in cell wall mechanics, cell elongation, and the axial growth of various vegetative and reproductive organs, as the loss of At KINESIN-4C greatly enhanced the defects caused by a null mutation at the KINESIN-4A/FRA 1 locus. The double mutant displayed a lack of cell wall softening at normal stages of rapid cell elongation. Furthermore, enhanced deposition of arabinose-containing carbohydrate was detected in the kinesin-4 mutants. Our findings established a connection between the Kinesin-4-based transport of cargoes containing non-cellulosic components along cortical microtubules and cell wall mechanics and cell elongation in flowering plants.

关键词： cytoskeleton cytoskeleton-cell wall interactions leaf/vegetative development Arabidopsis cotton

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The newly identified augmin complex is required in the assembly of the acentrosomal microtubule assay in higher plants

The newly identified augmin complex is required in the assem...

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从植物科学到农业发展——2012全国植物生物学大会

作者： yuh-ru julie lee Department of Plant Biology University of CaliforniaDavisCalifornia

In higher plant cells,microtubules(MTs) are nucleated and organized in a centrosome-independent *** is unclear whether augmin-dependent mechanisms underlie spindle MT organization in plant cells as they do in animal *** AUGMIN subunit3(AUG3),which encodes a homolog of animal dimy-tubulin 3/human augmin-like complex,subunit 3,was disrupted in Arabidopsis thaliana,gametogenesis frequently failed due to defects in cell *** with the controlmicrospores,which formed bipolar spindles at the cell periphery,the mutant cells often formed peripheral half spindlesthat only attached to condensed chromosomes or formed elongated spindles with unfocused interior *** addition,defective cells exhibited disorganized phragmoplast MT arrays,which caused aborted *** resulting pollengrains were either shrunken or contained two nuclei in an undivided ***3 was localized along MTs in thespindle and phragmoplast,and its signal was pronounced in anaphase spindle *** AUG3-green fluorescent proteinfusion exhibited a dynamic distribution pattern, similar to that of the y-tubulin complex *** usingrobustimmunoprecipitation and mass spectrometry,we further identified the entire plant augmin complex that contains eight *** them,AUG1 to AUG6 share low sequencesimilarity with their animal counterparts,but AUG7 and AUG8 share homology only with proteins of plant *** conclude that assembly ofMT arrays during plant mitosis depends on the augmin complex,which includes two plant-specific subunits.

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