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Formins:Bringing new insights to the organization of actin cytoskeleton

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Chinese Science Bulletin 2006年第24期51卷 2937-2943页

作者： GUO Chunqing ren haiyun Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education and College of Life Science Beijing Normal University Beijing 100875 China

The actin cytoskeleton is an important component of eukaryotic cell cytoskeleton and is temporally and spatially controlled by a series of actin binding proteins (ABPs). Among ABPs, formin family proteins have attracted much attention as they can nucleate unbranched actin filament from the profilin bound actin pool in vivo. In recent years, a number of formin family members from different organisms have been reported, and their characteristics are known more clearly, although some questions are still to be clarified. Here, we summarize the structures, func-tions and nucleation mechanisms of different formin family proteins, intending to compare them and give some new clues to the study of formins.

关键词： Formins蛋白成蛋白肌动蛋白细胞骨架肌动蛋白结合蛋白

Polymerization of fluorescent analogue of plant actin in vitro and in vivo

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Chinese Science Bulletin 2000年第17期45卷 1583-1586页

作者： ren haiyun YUAN Ming Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life SciencesBeijing Normal UniversityBeijing 100875China Key Laboratory of Plant Physiology and Biochemistry of Ministry of Agriculture Department of Plant SciencesCollege of Biological SciencesChina Agricultural UniversityBeijing 100094China

Maize pollen actin has been labeled with Oregon Green 488 iodoacetamide. A yield of 3 mg fluorescent actin analogue has been obtained from 10 mg of maize pollen actin, which is 99% in purity and the dye/protein ratio is 72%. In the presence of Mg2+ and K+, the fluorescent actin analogue polymerized into filaments in vitro. Green fluorescent filaments were observed when the fluorescent actin was introduced into living plant cells by microinjection, indicating that the fluorescent actin analogue functions similarly to the native actin.

关键词： fluorescent actin analogue actin filaments microinjection.

Actin Polymerization Mediated by AtFH5 Directs the Polarity Establishment and Vesicle Trafficking for Pollen Germination in Arabidopsis

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Molecular Plant 2018年第11期11卷 1389-1399页

作者： Chang Liu Yi Zhang haiyun ren Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life ScienceBeijing Normal UniversityBeijing 100875China

The process of pollen germination is crucial for flowering plant reproduction,but the mechanisms through which pollen grains establish polarity and select germination sites are not well *** this study,we report that a formin family protein,AtFH5,is localized to the vesicles and rotates ahead of Lifeact-mEGFPlabeled actin filaments during pollen *** translocation of AtFH5 to the plasma membrane initiates the assembly of a collar-like actin structure at the prospective germination site prior to germination. Genetic and pharmacological evidence further revealed an interdependent relationship between the mobility of AtFH5-1 abeled vesicles and the polymerization of actin filaments:vesicle-localized AtFH5 promotes actin assembly,and the polymerization and elongation of actin filaments,in turn,is essential for the mobility of AtFH5-1 abeled vesicles in pollen *** together,our work revealed a molecular mechanism underlying the polarity establishment and vesicle mobility during pollen germination.

关键词： Chinese cabbage Pollen germination Polarity establishment Vesicle trafficking Actin Formin

In vitro assembly of plant tubulin in the absence of microtubule-stabilizing reagents

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Chinese Science Bulletin 2000年第24期45卷 2258-2263页

作者： HUANG Shanjin ren haiyun YUAN Ming Key Laboratory of Plant Physiology and Biochemistry of Ministry of Agriculture Department of Plant SciencesCollege of Biological SciencesChina Agricultural UniversityBeijing 100094China Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life SciencesBeijing Normal UniversityBeijing 100875China

The assembly of microtubules is essential for physiological functions of microtubules. Addition of microtubule-stabilizing reagents or microtubule 'seeds' is usually necessary for plant tubulin assembly in vitro, which hinders the investigation of plant microtubule dynamics. In the present note, highly purified plant tubulins have been obtained from lily pollen, a non-microtubule-stabilizing reagent or microtubule 'seed' system for plant tubulin assembly has been established and the analysis of plant tubulin assembly performed. Experiment results showed that purified tubulin polymerized in vitro, and a typical microtubule structure was observed with electron microscopy. The kinetics curve of tubulin assembly exhibited typical 'parabola'. The presence of taxol significantly altered the character of plant tubulin assembly, including that abnormal microtubules were assembled and the critical concentration for plant tubulin assembly was decreased exceedingly from 3 mg/mL in the absence of taxol to 0.043 mg/

关键词： lily pollen tubulin microtubule tubulin assembly.

Actin Fringe Is Correlated with Tip Growth Velocity of Pollen Tubes

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Molecular Plant 2012年第5期5卷 1160-1162页

作者： Huaijian Dong Weike Pei ren haiyun Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education and College of Life Science Beijing Normal University Beijing 100875 China

Dear Editor, As an essential element for pollen tube tip growth, actJn cytoskeleton has been a focus for decades. Recently, both rapid-freeze and room-temperature fixation have revealed the presence of a subapical cortical F-actin （actin fringe） as a consistent feature of lily pollen tubes （Lovy-Wheeler et al., 2005）. The actin fringe consists of a palisade of short longitudinally oriented actin cables that start about 1-5~m behind the extreme apex and extend basally for an additional 5-10pm （Lovy-Wheeler et al., 2005）. MeanwhJJe, observations in live cells using Lifeact-mEGFP, an actin probe consisting of the first 17 amino acids from the budding yeast ABP-140 fused to GFP, which does not affect the dynamics of actin, compellingly support the presence of a cortical actin fringe in lilv pollen tubes ~Vidali et al.. 2009~.

关键词： F-肌动蛋白花粉管生长速度氨基酸组成必需元素顶端生长快速冷冻芽殖酵母

Dynamics and functions of the actin cytoskeleton during the plant cell cycle

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Chinese Science Bulletin 2011年第33期56卷 3504-3510页

作者： LIU PeiWei QI Ming XUE XiuHua ren haiyun Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life Science Beijing Normal UniversityBeijing 100875 China

In eukaryotic cells, the course of the cell cycle depends on correct cytoskeleton arrangement. The cell cycle consists of several phases, and in each of them the cytoskeleton has a unique structure and set of characteristics. The dynamics of the cytoskeleton together with its binding proteins greatly contribute to progression of the cell cycle. Here, we mainly review recent research on the dynamic distribution of the actin cytoskeleton and actin-binding proteins, and the mechanisms by which they affect the progression of the plant cell cycle.

关键词：肌动蛋白细胞骨架细胞周期植物肌动蛋白结合蛋白动力学真核细胞动态变化动态分布

At FH16, an Arabidopsis Type II Formin, Binds and Bundles both Microflaments and Microtubules, and Preferentially Binds to Microtubules

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Journal of Integrative Plant Biology 2013年第11期55卷 1002-1015页

作者： Jiaojiao Wang Yi Zhang Jing Wu Lei Meng haiyun ren The Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life Science Beijing Normal University

Formins are well-known regulators that participate in the organization of the actin cytoskeleton in organisms. The Arabidopsis thaliana L. genome encodes 21 formins, which can be divided into two distinct subfamilies. However, type II formins have to date been less well characterized. Here, we cloned a type II formin, AtFH16, and characterized its biochemical activities on actin and microtubule dynamics. The results show that the FH1 FH2 structure of AtFH16 cannot nucleate actin polymerization efficiently, but can bind and bundle microfilaments. AtFH16 FHIFH2 is also able to bind and bundle microtubules, and preferentially binds microtubules over microfilaments in vitro, in addition, AtFH16 FHIFH2 co-localizes with microtubules in onion epidermal cells, indicating a higher binding affinity of AtFH16 FHIFH2 for microtubules rather than microfilaments in vivo. In conclusion, AtFH16 is able to interact with both microfilaments and microtubules, suggesting that AtFH16 probably functions as a bifunctional protein, and may thus participate in plant cellular processes.

关键词： Actin filament Arabidopsis cytoskeleton formin microtubule.

A Processive Arabidopsis Formin Modulates Actin Filament Dynamics in Association with Profilin

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Molecular Plant 2016年第6期9卷 900-910页

作者： Sha Zhang Chang Liu Jiaojiao Wang Zhanhong ren Christopher J. Staiger haiyun ren Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life Science Beijing Normal University Beijing 100875 People'sRepublic of China Department of Biological Sciences Purdue University Hansen Life Sciences Research Building West Lafayette IN 47907-2064 USA

Formins are conserved regulators of actin cytoskeletal organization and dynamics that have been impli- cated to be important for cell division and cell polarity. The mechanism by which diverse formins regulate actin dynamics in plants is still not well understood. Using in vitro single-molecule imaging technology, we directly observed that the FH1-FH2 domain of an Arabidopsis thaliana formin, AtFH14, processively at- taches to the barbed end of actin filaments as a dimer and slows their elongation rate by 90%. The attach- ment persistence of FH1-FH2 is concentration dependent. Furthermore, by use of the triple-color total internal reflection fluorescence microscopy, we found that ABP29, a barbed-end capping protein, com- petes with FH1-FH2 at the filament barbed end, where its binding is mutually exclusive with AtFH14. In the presence of different plant profilin isoforms, FH1-FH2 enhances filament elongation rates from about 10 to 42 times. Filaments buckle when FH1-FH2 is anchored specifically to cover slides, further indicating that AtFH 14 moves processively on the elongating barbed end. At high concentration, AtFH 14 bundles actin filaments randomly into antiparallel or parallel spindle-like structures; however, the FH1-FH2-mediated bundles become thinner and longer in the presence of plant profilins. This is the direct demonstration of a processive formin from plants. Our results also illuminate the molecular mechanism of AtFH14 in regulating actin dynamics via association with profilin.

关键词： processive formin profilin capping protein actin bundle TIRF microscopy single molecule

Actin organization and regulation during pollen tube growth

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Frontiers in Biology 2011年第1期6卷 40-51页

作者： Xiuhua XUE Fei DU Jinsheng ZHU haiyun ren Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education College of Life SciencesBeijing Normal UniversityBeijing 100875China

Pollen is the male gametophyte of seed plants and its tube growth is essential for successful *** evidence has demonstrated that actin organization and regulation plays a central role in the process of its germination and polarized *** native structures and dynamics of actin are subtly modulated by many factors among which numerous actin binding proteins(ABPs)are the most direct and significant *** signals such as Ca^(2+),PIP_(2)(phosphatidylinositol-4,5-bis-phosphate)and GTPases can also indirectly act on actin organization through several *** such elaborate regulation,actin structures show dynamically continuous modulation to adapt to the in vivo biologic functions to mediate secretory vesicle transportation and fusion,which lead to normal growth of the pollen *** encouraging progress has been made in the connection between actin regulation and pollen tube growth in recent *** this review,we summarize different factors that affect actin organization in pollen tube growth and highlight relative research progress.

关键词： pollen tube tip growth actin organization actin-binding protein signaling pathway