Coupled, circumferential motions of the cell wall synthesis machinery and MreB filaments in B. subtilis

Rod-shaped bacteria elongate by the action of cell wall synthesis complexes linked to underlying dynamic MreB filaments. To understand how the movements of these filaments relate to cell wall synthesis, we characterized the dynamics of MreB and the cell wall elongation machinery using high-precision particle tracking in Bacillus subtilis. We found that MreB and the elongation machinery moved circumferentially around the cell, perpendicular to its length, with nearby synthesis complexes and MreB filaments moving independently in both directions. Inhibition of cell wall synthesis by various methods blocked the movement of MreB. Thus, bacteria elongate by the uncoordinated, circumferential movements of synthetic complexes that insert radial hoops of new peptidoglycan during their transit, possibly driving the motion of the underlying MreB filaments.     

Cryo-electron tomography reveals the cytoskeletal structure of Spiroplasma melliferum

Evidence has accumulated recently that not only eukaryotes but also bacteria can have a cytoskeleton. We used cryo-electron tomography to study the three-dimensional structure of Spiroplasma melliferum cells in a close-to-native state at approximately 4-nanometer resolution. We showed that these cells possess two types of filaments arranged in three parallel ribbons underneath the cell membrane. These two filamentous structures are built of the fibril protein and possibly the actin-like protein MreB. On the basis of our structural data, we could model the motility modes of these cells and explain how helical Mollicutes can propel themselves by means of coordinated length changes of their cytoskeletal ribbons.     

MreB在鱼腥蓝细菌PCC 7120细胞分裂过程中的功能

为研究Anabaena sp. PCC 7120中的细胞形态决定蛋白MreB的亚细胞定位和功能,构建了由mreB自身启动子驱动的mreB-gfp融合载体,通过接合转移的方法将其转化到野生型Anabaena sp. PCC 7120中,获得绿色荧光蛋白标记的MreB的鱼腥蓝细菌菌株,同时用MreB的抑制剂A22处理菌株。结果显示,MreB的亚细胞定位随着细胞周期的进程而发生变化,A22处理可导致MreB无法正确聚合定位。     

A trypanosome structure involved in transmitting cytoplasmic information during cell division

African trypanosomes are protozoan parasites that cause sleeping sickness in humans through a tsetse fly vector. The procyclic form of Trypanosoma brucei has a single, attached flagellum that describes a helical path along the cell from posterior to anterior. During division, a specific flagellum-flagellum connection is elaborated between the new and old flagellum. This connector was present only during cell duplication and was found to be involved in the replication of the helical cell pattern and polarity. This finding implicates the concept of cytotaxis in cell morphogenesis in trypanosomes.     

美洲黑杨次生木质部导管分化进程的超微结构分析

利用常规电镜技术,观察研究了美洲黑杨Populus deltoides次生木质部导管分化过程的超微结构变化。结果表明：美洲黑杨导管形态的建成可划分为初生壁的延展、次生壁的构建与穿孔板的形成等3个时期。初生壁的延展是导管分化的初始阶段,导管细胞高度液泡化,细胞质及其细胞器贴壁分布。次生壁的构建是导管分化的关键阶段,次生壁物质的沉积在导管液泡膜破裂之前即已开始,此时导管分子内细胞器结构清晰,其中高尔基体及其分泌小泡最为丰富,说明高尔基体与次生壁物质的合成及运输密切相关;导管分子液泡膜裂解后,次生壁物质沉积极为迅速,伴随次生壁物质的合成,导管分子细胞质解体,细胞核染色质凝聚并边缘化,表现出程序化细胞死亡（PCD）的典型特征。穿孔板的形成是导管分化的终极阶段,在导管次生壁形成时,相邻导管分子的端壁上不发生壁物质的积累,而且在次生壁构建后期,端壁上的壁物质降解,最后残余的端壁断裂形成穿孔板。美洲黑杨次生木质部导管的分化阶段彼此相继有序地进行,其中次生壁构建的启动是导管分子不可逆分化的临界期,随后的分化阶段是一种典型的PCD过程。     

Magnetosomes are cell membrane invaginations organized by the actin-like protein MamK

Magnetosomes are membranous bacterial organelles sharing many features of eukaryotic organelles. Using electron cryotomography, we found that magnetosomes are invaginations of the cell membrane flanked by a network of cytoskeletal filaments. The filaments appeared to be composed of MamK, a homolog of the bacterial actin-like protein MreB, which formed filaments in vivo. In a mamK deletion strain, the magnetosome-associated cytoskeleton was absent and individual magnetosomes were no longer organized into chains. Thus, it seems that prokaryotes can use cytoskeletal filaments to position organelles within the cell.     

Rotational movement of the formin mDia1 along the double helical strand of an actin filament

Formin homology proteins (formins) elongate actin filaments (F-actin) by continuously associating with filament tips, potentially harnessing actin-generated pushing forces. During this processive elongation, formins are predicted to rotate along the axis of the double helical F-actin structure (referred to here as helical rotation), although this has not yet been definitively shown. We demonstrated helical rotation of the formin mDia1 by single-molecule fluorescence polarization (FL(P)). FL(P) of labeled F-actin, both elongating and depolymerizing from immobilized mDia1, oscillated with a periodicity corresponding to that of the F-actin long-pitch helix, and this was not altered by actin-bound nucleotides or the actin-binding protein profilin. Thus, helical rotation is an intrinsic property of formins. To harness pushing forces from growing F-actin, formins must be anchored flexibly to cell structures.     

螺原体检测方法概述

螺原体（spiroplasma）是一种个体极小（直径约为100～250nm）,基本形态为螺旋形的无细胞壁的原核微生物。1973年建立了螺原体属（Spiroplasma）。与一般细菌相比,螺原体的研究方法,特别是螺原体的检测方法比较特殊。对4种螺原体检测方法,即分离培养、显微镜检查、血清学检测以及分子生物学检测方法进行了概述。     

Fungal morphogenesis: cell wall construction in Mucor rouxii

An autoradiographic study revealed two different patterns of cell wall constructioni associated with two types of morphogenesis in Mucor rouxii. In hyphae, the cell wall was preferentially synthesized in the apical region; these cylindrical walls seemed to be generated by a sharply descending gradient of wall synthesis radiating from the apex. In spherical cells (germinating spores, yeast cells), wall formation occurred largely, if not entirely, in uniformly dispersed fashion over the entire cell periphery.     

Temporal frequency of subthreshold oscillations scales with entorhinal grid cell field spacing

Grid cells in layer II of rat entorhinal cortex fire to spatial locations in a repeating hexagonal grid, with smaller spacing between grid fields for neurons in more dorsal anatomical locations. Data from in vitro whole-cell patch recordings showed differences in frequency of subthreshold membrane potential oscillations in entorhinal neurons that correspond to different positions along the dorsal-to-ventral axis, supporting a model of physiological mechanisms for grid cell responses.     

BRITTLE CULM16(BRITTLE NODE) is required for the formation of secondary cell walls in rice nodes

Plant cell walls constitute the skeletal structures of plant bodies,and thus confer lodging resistance for grain crops.While the basic cell wall synthesis machinery is relatively well established now,our understanding of how the process is regulated remains limited and fragmented.In this study,we report the identification and characterization of the novel rice(Oryza sativa L.)brittle culm16(brittle node;bc16)mutant.The brittle node phenotype of the bc16 mutant appears exclusively at nodes,and resembles the previously reported bc5 mutant.Combined histochemical staining and electron microscopy assays revealed that in the bc16 mutant,the secondary cell wall formation and thickening of node sclerenchyma tissues are seriously affected after heading.Furthermore,cell wall composition assays revealed that the bc16 mutation led to a significant reduction in cellulose and lignin contents.Using a map-based cloning approach,the bc16 locus is mapped to an approximately 1.7-Mb region of chromosome 4.Together,our findings strengthen evidence for discretely spatial differences in the secondary cell wall formation within plant bodies.     

Helical filaments produced by a Mycoplasma-like organism associated with corn stunt disease

Mycoplasma-like bodies with helical filaments were seen by phase contrast microscopy in juice expressed from tissues of plants infected with corn stunt agent. Each filament is bounded by a "unit membrane" and no cell wall, sheath, envelope, or second membrane has yet been discerned by electron microscopy. The association of these filaments with development of disease, their occurrence in phloem cells as seen by both freeze-etching and thin-section electron microscopy, the diagnosis of infection based on their presence in plants without symptoms, and their absence in noninfected corn are consistent with the hypothesis that these unusual filaments are formed by the mycoplasma-like organism presumed to be the corn stunt agent.     

棉花纤维品质改良相关基因研究进展

棉纤维是优良的、使用最为广泛的天然纤维。随着人们生活水平的提高,对天然纯棉织物的需求不断增加,同时对品质的要求也愈来愈高。因此,提高棉纤维产量和品质成为当前棉花遗传育种的重要目标,对棉纤维发育相关基因的克隆与功能研究是实现这一目标的重要基础。棉纤维发育由4个明显但又重叠的时期组成,包括纤维细胞的起始、伸长(初生壁合成)、次生壁合成和脱水成熟。起始是影响纤维细胞数量的重要时期,而纤维长度和强度的决定发生在次生壁合成期和脱水成熟期。棉纤维发育是一个复杂而有序的过程,由大量的基因参与调控。目前,已经有一些在棉纤维发育过程中发挥重要作用的基因被报道,包括各种转录因子、参与激素代谢基因、编码细胞壁蛋白和细胞骨架蛋白基因、活性氧代谢相关基因、以及参与糖和脂类代谢的基因等。文中对已报道的这些与棉花纤维发育相关基因的克隆和功能分析进行了系统总结,以期为棉花纤维发育及品质改良研究提供参考。     

Substructure of certain cytoplasmic microtubules: an electron microscopic study

egatively stained cytoplasmic microtubules of lung-fluke sperm show a helical structure that is not found in peripheral doublet tubules of axial units. In transverse sections, the wall of such microtubules appears to comprise about eight subunits.     

针叶材管胞细胞壁不同壁层的纵向弹性模量和硬度

该文利用原位成像纳米压痕技术研究了针叶材管胞细胞壁不同壁层之间在纵向弹性模量 (MOE) 和硬度方面的差异．结果表明,当压针从细胞壁的纵向压入时,细胞壁变形机制以塑性变形为主;细胞壁纵向弹性模量和硬度在细胞壁厚方向的分布不均匀．S3层与细胞腔交界处、S1层与复合胞间层(CML)交界处的弹性模量和硬度均明显小于次生壁(SW)S2层．此外,相邻管胞次生壁S2层之间的弹性模量和硬度也存在一定差异．ANOVA分析表明,杉木管胞、马尾松管胞次生壁的弹性模量均显著大于复合胞间层,但两者之间的硬度差异不显著．虽然马尾松成熟材管胞次生壁的纵向弹性模量和硬度均大于幼龄材,但差异的程度不同．弹性模量前者比后者约大40％,但硬度只大13％．管胞次生壁和复合胞间层的弹性模量和硬度之间均存在较显著的正线性相关关系．      

基于根茎叶定量概化模型的植被区沿程波高衰减机制试验研究

  目的  红树林作为一种新型生态式的护岸工具，其消波效应已成为海岸工程研究热点。本文对红树林根、茎、叶不同组成部分进行定量研究，进一步探究红树林对波浪的消减效应。  方法  采取定量概化的根茎叶植物模型，通过波浪水槽试验探究了红树林植被区沿程波高衰减机制。将Kobayashi等提出的理论模型计算所得的植被区沿程波高理论值与试验值进行对比。  结果  研究发现，由Kobayashi等提出的理论模型计算所得的试验波高衰减系数的范围分别为0.02 ~ 0.47。当入射波波长与植被区沿流向长度之比趋于1时，消波强度最弱。同时波浪在植被区衰减过程中出现了边界效应及波浪跃增现象，波高增量比的范围为0 ~ 13%。当波长与植被区沿流向长度之比在1.0 ~ 1.2之间，且淹没度等于1.0时，跃增高度随叶的分布密度的减小而增大，随根的分布密度的减小而减小。而当淹没度等于0.8时，减小根或叶的分布密度，跃增高度均减小。  结论  根和叶的分布密度、淹没度以及波长与植被区沿流向长度之比均会对植被区沿程波高衰减规律产生不同程度的影响。      

紫花苜蓿MsNST的克隆及对木质素与纤维素合成的功能分析

【目的】木质素和纤维素是植物次生细胞壁的主要组成成分,是影响牧草消化率和品质的主要因素之一。紫花苜蓿作为高蛋白饲草,是奶牛等草食家畜优质饲草的主要来源。因此,苜蓿木质素和纤维素合成机制一直是受到关注的研究热点。模式植物拟南芥NAC家族的NST转录因子（NAC secondary wall thicking promoting factor）调控次生细胞壁的合成,但紫花苜蓿NST的功能与调控机制尚不明确。本研究通过分析紫花苜蓿NST的表达模式,及在拟南芥与苜蓿中过表达揭示其对木质素和纤维素合成的影响。【方法】通过同源克隆获得MsNST CDs序列,并对该基因进行生物信息学分析。利用qRT-PCR检测该基因受赤霉素（GA3）,水杨酸（SA）和多效唑（PCB）诱导后的表达模式。通过转基因植株中过表达MsNST,研究其对木质素和纤维素含量及其合成相关基因的影响。【结果】克隆MsNST 的CDs序列,最大开放阅读框为945bp,编码314个氨基酸。生物信息学分析表明,MsNST主要由无规则卷曲组成（60.83 %）;三级结构预测显示,MsNST以同源二聚体的形式有效的促进蛋白质间的相互作用。进化树分析表明,单子叶和双子叶分为两个分枝,暗示存在一定程度的进化,而MsNST与蒺藜苜蓿和大豆的NST属于双子叶植物分枝中的亚分枝,表明豆科植物间亲缘关系较近。MsNST与拟南芥NST1-3的氨基酸序列相似性较高（49%—55.9%）,并含有NAC转录因子的5个保守结构域。qRT-PCR分析表明,MsNST受GA3、SA和PCB的诱导表达,相对表达水平（12h）分别是对照的2.19、3.67和3.65倍。过表达MsNST导致拟南芥下胚轴缩短,转基因拟南芥半矮化,花序茎束间细胞壁纤维增厚,细胞壁结晶纤维素（13%）、总糖（7%）和木质素（11.7%）含量增加。通过分析木质素和纤维素合成相关基因表达水平发现,拟南芥和苜蓿中过表达MsNST均能激活木质素合成关键基因（PAL,4CL等）及纤维素合酶复合体亚基CesA家族基因的表达。【结论】MsNST受外源激素GA3、SA和PCB诱导表达。转基因植物中过表达MsNST可激活次生壁木质素和纤维素合成相关基因的表达,同时茎束间纤维细胞壁增厚,细胞壁结晶纤维素、总糖和木质素含量增加,暗示MsNST对次生细胞壁木质素和纤维素的合成有重要的调节作用。     

Mechanism of two classes of cancer mutations in the phosphoinositide 3-kinase catalytic subunit

Many human cancers involve up-regulation of the phosphoinositide 3-kinase PI3Kalpha, with oncogenic mutations identified in both the p110alpha catalytic and the p85alpha regulatory subunits. We used crystallographic and biochemical approaches to gain insight into activating mutations in two noncatalytic p110alpha domains-the adaptor-binding and the helical domains. A structure of the adaptor-binding domain of p110alpha in a complex with the p85alpha inter-Src homology 2 (inter-SH2) domain shows that oncogenic mutations in the adaptor-binding domain are not at the inter-SH2 interface but in a polar surface patch that is a plausible docking site for other domains in the holo p110/p85 complex. We also examined helical domain mutations and found that the Glu545 to Lys545 (E545K) oncogenic mutant disrupts an inhibitory charge-charge interaction with the p85 N-terminal SH2 domain. These studies extend our understanding of the architecture of PI3Ks and provide insight into how two classes of mutations that cause a gain in function can lead to cancer.     

Metals and DNA: molecular left-handed complements

Chiral metal complexes provide unique molecular probes for DNA. Chiral reagents that "recognize" different local structures along the DNA strand have been designed by a process in which the asymmetry in shape and size of the complex is matched to that of the DNA helical groove. As a result, the chiral metal complexes provide very sensitive probes for local helical structure, both left- and right-handed. Direct coordination of chiral complexes to the DNA bases adds an element of sequence selectivity to the probe design. With a suitable reactive metal center, reagents that target chemically specific sites along the strand may be developed. One such chiral reagent, which cleaves left-handed DNA sites with photoactivation, has been useful in mapping this distinct conformation and examining its biological role. The conformation-specific molecular cleaver, much like a DNA-binding enzyme, recognizes and reacts at discrete sites along the DNA strand. These site-specific chiral metal complexes provide exciting new tools for probing the local variations in DNA structure and its role in the regulation of gene expression.     

植物与病原物互作中的活性氧代谢及其作用

介绍了活性氧的种类、代谢途径和测定方法,以及植物人存在的抗氧化机制、植物与病原物互作中活性氧的产生及遗传基础,重点对活性氧在植物与病原物互作中抗病信号转导、抗菌作用、膜脂氧化、细胞壁强化、植保素合成和过敏性细胞死亡等方面进行了综述。