共查询到20条相似文献,搜索用时 15 毫秒
1.
The plant actin depolymerizing factor (ADF) binds to both monomeric and filamentous actin, and is directly involved in the depolymerization of actin filaments. To better understand the actin binding sites of the Arabidopsis thaliana L. AtADF1, we generated mutants of AtADF1 and investigated their functions in vitro and in vivo. Analysis of mutants harboring amino acid substitutions revealed that charged residues (Arg98 and Lys100) located at the α‐helix 3 and forming an actin binding site together with the N‐terminus are essential for both G‐ and F‐actin binding. The basic residues on the β‐strand 5 (K82/A) and the α‐helix 4 (R135/A, R137/A) form another actin binding site that is important for F‐actin binding. Using transient expression of CFP‐tagged AtADF1 mutant proteins in onion (Allium cepa) peel epidermal cells and transgenic Arabidopsis thaliana L. plants overexpressing these mutants, we analyzed how these mutant proteins regulate actin organization and affect seedling growth. Our results show that the ADF mutants with a lower affinity for actin filament binding can still be functional, unless the affinity for actin monomers is also affected. The G‐actin binding activity of the ADF plays an essential role in actin binding, depolymerization of actin polymers, and therefore in the control of actin organization. 相似文献
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Shigehiro Hirano Tomikazu Nishio Tatsuro Ito 《Bioscience, biotechnology, and biochemistry》2013,77(10):1963-1967
The action of P2O5 in DMSO on methyl α-d-glucopyranoside, sucrose and trehalose afforded nondializable, phosphorylated glycans in 6~34% yields. Polysucrose has a molecular weight of ~9,500. The synthetic glycans consist of carbohydrate (46~59%) and P (11.4~13.1%) and show reducing sugar values (5.0~30.8%). The alkaline hydrolysis of polysucrose was accompanied with a depolymerization and afforded sugar phosphates and oligosaccharides. The periodate oxidation gave formic acid (0.15~0.34 mole) and formaldehyde (0.07~0.17 mole/monosaccharide residue). The methylation study indicated their variously branched structures. 2,3,4,6-Tetra-O-methyl-d-glucose was found in only 0.7 ~3.2% yields, and this is in agreement with their weak precipitation reactions with Con A. It is considered that the glycans are produced from nonreducing mono- and oligosaccharides by dehydration, transglycosidation and esterification with phosphate. 相似文献
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M H Buc-Caron D Lamblin O Kellermann 《Biology of the cell / under the auspices of the European Cell Biology Organization》1989,65(2):195-198
The visceral endoderm of the mouse embryo is a polarized epithelium which has recently been shown to express villin, a major actin binding component of absorptive epitheliums. We report here that villin is induced during differentiation of aggregates of the mouse embryonal carcinoma F9, an in vitro system widely used to study extraembryonic endoderm differentiation. Identical results were obtained with a variant of F9 which carries an immortalizing vector. Villin is coexpressed with F-actin and with alpha-foetoprotein, in most of the visceral endoderm-like cells lining the aggregates. This system is potentially useful to study (i) the induction of villin expression and (ii) the establishment of polarity in the visceral endoderm epithelium. 相似文献
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植物肌动蛋白异型体研究进展 总被引:7,自引:0,他引:7
肌动蛋白在真核生物中广泛存在,由肌动蛋白参与形成的动态微丝骨架系统是细胞生命活动的基础。在植物细胞中,肌动蛋白由多基因家族编码,从而产生了多种肌动蛋白异型体。本文综述了拟南芥肌动蛋白异型体的分类、体内分布与功能,详细介绍了豌豆卷须肌动蛋白异型体的研究现状,并讨论了该领域的研究方向。 相似文献
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Mariapina D'Onofrio Alejandro Giorgetti Paola Dominici Alessandra Astegno 《Protein science : a publication of the Protein Society》2016,25(8):1461-1471
In addition to the well‐known Ca2+ sensor calmodulin, plants possess many calmodulin‐like proteins (CMLs) that are predicted to have specific roles in the cell. Herein, we described the biochemical and biophysical characterization of recombinant Arabidopsis thaliana CML14. We applied isothermal titration calorimetry to analyze the energetics of Ca2+ and Mg2+ binding to CML14, and nuclear magnetic resonance spectroscopy, together with intrinsic and ANS‐based fluorescence, to evaluate the structural effects of metal binding and metal‐induced conformational changes. Furthermore, differential scanning calorimetry and limited proteolysis were used to characterize protein thermal and local stability. Our data demonstrate that CML14 binds one Ca2+ ion with micromolar affinity (Kd ~ 12 µM) and the presence of 10 mM Mg2+ decreases the Ca2+ affinity by ~5‐fold. Although binding of Ca2+ to CML14 increases protein stability, it does not result in a more hydrophobic protein surface and does not induce the large conformational rearrangement typical of Ca2+ sensors, but causes only localized structural changes in the unique functional EF‐hand. Our data, together with a molecular modelling prediction, provide interesting insights into the biochemical properties of Arabidopsis CML14 and may be useful to direct additional studies aimed at understanding its physiological role. 相似文献
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Hui Su Ting Wang Huaijian Dong Haiyun Ren 《植物学报(英文版)》2007,49(8):1183-1191
The villin/gelsolin/fragmin superfamily is a conserved Ca^2+-dependent family of actin-regulating proteins that is widely present both in mammalian and non-mammalian organisms. They have traditionally been characterized by the same core of three or six tandem gelsolin subdomains. The study in vertebrates and lower eukaryotic cells has revealed that the villin/gelsolin/fragmin superfamily of proteins has versatile functions including severing, capping, nucleating or bundling actin filaments. In plants, encouraging progress has been made in this field of research in recent years. This review will summarize the identified plant homologs of villin/gelsolin/fragmin superfamily, thus providing a basis for reflection on their biochemical activities and functions in plants. 相似文献
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拟南芥雄性不育突变体ms1142的遗传定位与功能分析 总被引:1,自引:0,他引:1
经EMS诱变野生型拟南芥(Arabidopsis thaliana)群体筛选得到一株雄性不育突变体ms1142, 突变体的果荚短小, 不含种子。细胞学观察和扫描电镜结果表明, 突变体花药发育过程中, 花药中小孢子外壁异常、破裂, 最后没有花粉形成。遗传分析表明, 该突变体为隐性单核基因突变所致; 利用图位克隆的方法将MS1142基因定位于第1条染色体的BAC克隆F16P17上44 kb区间内, 目前尚未见该区间内有雄性不育基因的报道。以上结果结合生物信息学分析表明, MS1142是一个新的调控花药发育的关键基因。该工作为花药发育关键基因MS1142的克隆及功能分析奠定了基础。 相似文献
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Liliya Vugmeyster Tien Do Dmitry Ostrovsky Riqianq Fu 《Protein science : a publication of the Protein Society》2014,23(2):145-156
Thermostable villin headpiece protein (HP67) consists of the N‐terminal subdomain (residues 10–41) and the autonomously folding C‐terminal subdomain (residues 42–76) which pack against each other to form a structure with a unified hydrophobic core. The X‐ray structures of the isolated C‐terminal subdomain (HP36) and its counterpart in HP67 are very similar for the hydrophobic core residues. However, fine rearrangements of the free energy landscape are expected to occur because of the interactions between the two subdomains. We detect and characterize these changes by comparing the µs‐ms time scale dynamics of the methyl‐bearing side chains in isolated HP36 and in HP67. Specifically, we probe three hydrophobic side chains at the interface of the two subdomains (L42, V50, and L75) as well as at two residues far from the interface (L61 and L69). Solid‐state deuteron NMR techniques are combined with computational modeling for the detailed characterization of motional modes in terms of their kinetic and thermodynamic parameters. The effect of interdomain interactions on side chain dynamics is seen for all residues but L75. Thus, changes in dynamics because of subdomain interactions are not confined to the site of perturbation. One of the main results is a two‐ to threefold increase in the value of the activation energies for the rotameric mode of motions in HP67 compared with HP36. Detailed analysis of configurational entropies and heat capacities complement the kinetic view of the degree of the disorder in the folded state. 相似文献
10.
Xiaoyue Chen Xuanhao Xu Yuna Sun Jingwen Zhou Yuanyuan Ma Liming Yan Zhiyong Lou 《Acta Crystallographica. Section F, Structural Biology Communications》2012,68(1):69-72
Plant‐specific dynamin‐related proteins play crucial roles in cell‐plate formation, endocytosis or exocytosis, protein sorting to the vacuole and plasma membrane and the division of mitochondria and chloroplasts. In order to determine the crystal structure and thus to obtain a better understanding of the biological functions and mechanisms of dynamin‐related proteins in plant cells, the GTPase domain of Arabidopsis thaliana dynamin‐related protein 1A (AtDRP1A) fused to its GTPase effector domain (GED) was crystallized in a nucleotide‐associated form using polyethylene glycol 3350 as precipitant. The hexagonal crystals (space group P61) had unit‐cell parameters a = b = 146.2, c = 204.3 Å, and diffraction data were collected to 3.6 Å resolution using synchrotron radiation. Four molecules, comprising two functional dimers, are assumed per asymmetric unit, corresponding to a Matthews coefficient of 3.9 Å3 Da−1 according to the molecular weight of 39 kDa. 相似文献
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Christel Verboven Ilse Bogaerts Etienne Waelkens Anja Rabijns Hugo Van Baelen Roger Bouillon Camiel De Ranter 《Acta Crystallographica. Section D, Structural Biology》2003,59(2):263-273
The multifunctional vitamin D binding protein (DBP) is an actin‐sequestering protein present in blood. The crystal structure of the actin–DBP complex was determined at 2.4 Å resolution. DBP binds to actin subdomains 1 and 3 and occludes the cleft at the interface between these subdomains. Most remarkably, DBP demonstrates an unusually large actin‐binding interface, far exceeding the binding‐interface areas reported for other actin‐binding proteins such as profilin, DNase I and gelsolin. The fast‐growing side of actin monomers is blocked completely through a perfect structural fit with DBP, demonstrating how DBP effectively interferes with actin‐filament formation. It establishes DBP as the hitherto best actin‐sequestering protein and highlights its key role in suppressing and preventing extracellular actin polymerization. 相似文献
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Jäger K Fábián A Tompa G Deák C Höhn M Olmedilla A Barnabás B Papp I 《Plant biology (Stuttgart, Germany)》2011,13(1):78-84
This paper provides a detailed phenotypic analysis of the abscisic acid (ABA) hypersensitive Cap Binding Protein 20 (cbp20) mutant. Some hitherto undescribed changes were found in the tissue structure and epidermal morphology of this mutant. These include more and smaller cells in the epidermis, a thicker cuticle and more frequent occurrence of trichomes on leaf surfaces. Some of these traits may contribute to the physiological processes responsible for the water-saving behaviour of the mutant. Abnormal spatial patterns between stomatal pore complexes were also found on various organs of the mutant. All these observations indicate profoundly disturbed development of epidermal tissue in the cbp20 mutant, which has not previously been reported for this class of mutants. A potential connection between the new phenotypes and disturbed miRNA metabolism and mRNA splicing of the mutant is discussed. 相似文献
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Ming Chang Zhankun Li Shanjin Huang 《The Plant journal : for cell and molecular biology》2017,92(3):509-519
Dynamic assembly and disassembly of the actin cytoskeleton has been implicated in the regulation of pollen germination and subsequent tube growth. It is widely accepted that actin filaments are arrayed into distinct structures within different regions of the pollen tube. Maintenance of the equilibrium between monomeric globular actin (G‐actin) and filamentous actin (F‐actin) is crucial for actin assembly and array construction, and the local concentration of G‐actin thus directly impacts actin assembly. The localization and dynamics of G‐actin in the pollen tube, however, remain to be determined conclusively. To address this question, we created a series of fusion proteins between green fluorescent protein (GFP) and the Arabidopsis reproductive actin ACT11. Expression of a fusion protein with GFP inserted after methionine at position 49 within the DNase I‐binding loop of ACT11 (GFPMet49–ACT11) rescued the phenotypes in act11 mutants. Consistent with the notion that the majority of actin is in its monomeric form, GFPMet49–ACT11 and GFP fusion proteins of four other reproductive actins generated with the same strategy do not obviously label filamentous structures. In further support of the functionality of these fusion proteins, we found that they can be incorporated into filamentous structures in jasplakinolide (Jasp)‐treated pollen tubes. Careful observations showed that G‐actin is distributed uniformly in the pollen tube and is rapidly redistributed via cytoplasmic streaming during pollen tube growth. Our study suggests that G‐actin is readily available in the cytoplasm to support continuous actin polymerization during rapid pollen tube growth. 相似文献
16.
A new class of proteins capable of binding transition metals 总被引:1,自引:0,他引:1
Dykema PE Sipes PR Marie A Biermann BJ Crowell DN Randall SK 《Plant molecular biology》1999,41(1):139-150
Ion uptake, transport, and sequestration are essential to meet the nutritional requirements for plant growth and development. Furthermore, regulation of these processes is critical for plants to tolerate toxic levels of ions. The examination of isoprenylated proteins encoded by Arabidopsis thaliana and Glycine max cDNAs revealed a unique family of proteins containing putative metal-binding motifs (the core sequence is M/LXCXXC). Here, we describe this new class of proteins, which are capable of being isoprenylated and binding transition metal ions. Members of this family contain consensus isoprenylation (CaaX) sites, which we demonstrate are efficiently isoprenylated in vitro. ATFP3, a representative of the Arabidopsis family, was expressed in Escherichia coli and examined for metal-binding activity in vitro. Analysis of the interaction of ATFP3 with metal-chelating columns (IMAC) suggested that it binds to Cu2+, Ni2+, or Zn2+. To test whether proteins with these characteristics are present in other plant species, tobacco BY2 cells were labeled in vivo with [14C]mevalonate and the resulting mevalonate-labeled proteins were tested for metal-binding activity. Several soluble, isoprenylated proteins which bound copper-IMAC columns were revealed. Consistent with a wide-spread distribution of these proteins in plants, their presence was observed in Arabidopsis, soybean, and tobacco. 相似文献
17.
Vladimir Sobolev Marvin Edelman Orly Dym Tamar Unger Shira Albeck Menny Kirma Gad Galili 《Acta Crystallographica. Section F, Structural Biology Communications》2013,69(2):84-89
Diaminopimelate aminotransferase (DAP‐AT) is an enzyme in the lysine‐biosynthesis pathway. Conversely, ALD1, a close homologue of DAP‐AT in plants, uses lysine as a substrate in vitro. Both proteins require pyridoxal‐5′‐phosphate (PLP) for their activity. The structure of ALD1 from the flowering plant Arabidopsis thaliana (AtALD1) was solved at a resolution of 2.3 Å. Comparison of AtALD1 with the previously solved structure of A. thaliana DAP‐AT (AtDAP‐AT) revealed similar interactions with PLP despite sequence differences within the PLP‐binding site. However, sequence differences between the binding site of AtDAP‐AT for malate, a purported mimic of substrate binding, and the corresponding site in AtALD1 led to different interactions. This suggests that either the substrate itself, or the substrate‐binding mode, differs in the two proteins, supporting the known in vitro findings. 相似文献
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Cloning and expression analysis of a gene that shows developmental regulation upon tuberization in potato 总被引:6,自引:0,他引:6
Jackson Stephen Gascón Jordi Carrera Esther Monte Elena Prat Salomé 《Plant molecular biology》1997,34(1):169-174
An Arabidopsis cDNA clone encoding a DNA-binding protein, RAP-1, was isolated by southwestern screening of an Escherichia coli cDNA expression library. The protein contains a bHLH DNA-binding domain and is homologous to R proteins, regulating anthocyanin biosynthesis. RAP-1 binds to the sequence CACNTG. It is encoded by a single gene, which is expressed to high levels in root and stem and to low levels in leaf and flower. No expression could be detected in siliques. Rap-1 does not correspond to one of the known loci involved in anthocyanin biosynthesis, since it is located at a different map position. In contrast to the maize R protein Lc, RAP-1 did not induce anthocyanin biosynthesis in pea cotyledons. Thus, RAP-1 is a novel member of the bHLH class of DNA-binding proteins. 相似文献
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