Directional switching of the kinesin Cin8 through motor coupling

Kinesin motor proteins are thought to move exclusively in either one or the other direction along microtubules. Proteins of the kinesin-5 family are tetrameric microtubule cross-linking motors important for cell division and differentiation in various organisms. Kinesin-5 motors are considered to be plus-end-directed. However, here we found that purified kinesin-5 Cin8 from budding yeast could behave as a bidirectional kinesin. On individual microtubules, single Cin8 motors were minus-end-directed motors, whereas they switched to plus-end-directed motility when working in a team of motors sliding antiparallel microtubules apart. This kinesin can thus change directionality of movement depending on whether it acts alone or in an ensemble.     

Differential regulation of dynein and kinesin motor proteins by tau

Dynein and kinesin motor proteins transport cellular cargoes toward opposite ends of microtubule tracks. In neurons, microtubules are abundantly decorated with microtubule-associated proteins (MAPs) such as tau. Motor proteins thus encounter MAPs frequently along their path. To determine the effects of tau on dynein and kinesin motility, we conducted single-molecule studies of motor proteins moving along tau-decorated microtubules. Dynein tended to reverse direction, whereas kinesin tended to detach at patches of bound tau. Kinesin was inhibited at about a tenth of the tau concentration that inhibited dynein, and the microtubule-binding domain of tau was sufficient to inhibit motor activity. The differential modulation of dynein and kinesin motility suggests that MAPs can spatially regulate the balance of microtubule-dependent axonal transport.     

Sustained microtubule treadmilling in Arabidopsis cortical arrays

Plant cells create highly structured microtubule arrays at the cell cortex without a central organizing center to anchor the microtubule ends. In vivo imaging of individual microtubules in Arabidopsis plants revealed that new microtubules are initiated at the cell cortex and exhibit dynamics at both ends. Polymerization-biased dynamic instability at one end and slow depolymerization at the other end result in sustained microtubule migration across the cell cortex by a hybrid treadmilling mechanism. This motility causes widespread microtubule repositioning and contributes to changes in array organization through microtubule reorientation and bundling.     

Molecular linkage underlying microtubule orientation toward cortical sites in yeast

Selective microtubule orientation toward spatially defined cortical sites is critical to polarized cellular processes as diverse as axon outgrowth and T cell cytotoxicity. In yeast, oriented cytoplasmic microtubules align the mitotic spindle between mother and bud. The cortical marker protein Kar9 localizes to the bud tip and is required for the orientation of microtubules toward this region. Here, we show that Kar9 directs microtubule orientation by acting through Bim1, a conserved microtubule-binding protein. Bim1 homolog EB1 was originally identified through its interaction with adenomatous polyposis coli (APC) tumor suppressor, raising the possibility that an APC-EB1 linkage orients microtubules in higher cells.     

Positioning of the mitotic spindle by a cortical-microtubule capture mechanism

Correct positioning of the mitotic spindle is critical for cell division and development. Spindle positioning involves a search-and-capture mechanism whereby dynamic microtubules find and then interact with specific sites on the submembrane cortex. Genetic, biochemical, and imaging experiments suggest a mechanism for cortical-microtubule capture. Bim1p, located at microtubule distal ends, bound Kar9p, a protein associated with the daughter cell cortex. Bim1p is the yeast ortholog of human EB1, a binding partner for the adenomatous polyposis coli tumor suppressor. EB1 family proteins may have a general role in linking the microtubule cytoskeleton to cortical polarity determinants.     

A gradient of sequence divergence in the human adult alpha-globin duplication units

The nucleotide sequences of the two 5'-homology blocks of human alpha-globin gene duplication units were determined. The sequence difference between the two blocks is essentially zero in the 5' portions, and increases gradually toward the 3' ends until it reaches a value of 18 percent. This gradient of sequence divergence is similar to the distribution of the frequencies of gene conversion along several loci in Ascobolus and yeast. Hot spots for initiation of gene correction processes appear to exist near the 5' ends of the human alpha-globin duplication units. The data provide the physical evidence for polar gene correction process in a mammalian genome.     

Dynamic instability of sheared microtubules observed by quasi-elastic light scattering

The kinetics of microtubule reassembly was studied in vitro by quasi-elastic light scattering (QELS). When microtubules assembled in the absence of microtubule-associated proteins (MAPs) were sheared, they rapidly depolymerized, recovered, and reassembled. The mean length of the recovered microtubules was the same as that observed just before shearing, implying that on average one fragment per original microtubule survived the fragmentation and recovery. When microtubules that contained 25 percent brain MAP were sheared, the fragments did not depolymerize extensively and the average length of the fragments decreased by a factor of 3 relative to the unsheared sample. The results support the dynamic instability model, which predicts that cellular microtubules are latently unstable structures protected on their ends by stabilizing caps.     

Cell cycle. Replication meets cohesion.

When a cell replicates its DNA during S phase of the cell cycle, the sister chromatid pairs must stick together like glue until they are separated to opposite ends of the cell (and hence into separate daughter cells) at anaphase. How the cell achieves this is still unclear but, as Takahashi and Yanagida explain in their Perspective, new findings in yeast have identified one molecule, Trf4p, that may be involved both in DNA replication and sister chromatid cohesion (Wang et al.).     

Direct measurements of sliding between outer doublet microtubules in swimming sperm flagella

The relative motion of 40-nanometer gold beads bound to the exposed outer doublet microtubules of demembranated sea urchin sperm flagella has been observed and photographed during adenosine triphosphate (ATP)-reactivated swimming. This direct demonstration and measure of sliding displacements between outer doublet microtubules in actively bending flagella verifies the original sliding microtubule model for ciliary bending that was established by electron microscopy of fixed cilia and provides a new, functional measure for the diameter of the flagellar axoneme of 132 +/- 8 nanometers.     

Pot1, the putative telomere end-binding protein in fission yeast and humans

Telomere proteins from ciliated protozoa bind to the single-stranded G-rich DNA extensions at the ends of macronuclear chromosomes. We have now identified homologous proteins in fission yeast and in humans. These Pot1 (protection of telomeres) proteins each bind the G-rich strand of their own telomeric repeat sequence, consistent with a direct role in protecting chromosome ends. Deletion of the fission yeast pot1+ gene has an immediate effect on chromosome stability, causing rapid loss of telomeric DNA and chromosome circularization. It now appears that the protein that caps the ends of chromosomes is widely dispersed throughout the eukaryotic kingdom.     

Motor cortical encoding of serial order in a context-recall task

The neural encoding of serial order was studied in the motor cortex of monkeys performing a context-recall memory scanning task. Up to five visual stimuli were presented successively on a circle (list presentation phase), and then one of them (test stimulus) changed color; the monkeys had to make a single motor response toward the stimulus that immediately followed the test stimulus in the list. Correct performance in this task depends on memorization of the serial order of the stimuli during their presentation. It was found that changes in neural activity during the list presentation phase reflected the serial order of the stimuli; the effect on cell activity of the serial order of stimuli during their presentation was at least as strong as the effect of motor direction on cell activity during the execution of the motor response. This establishes the serial order of stimuli in a motor task as an important determinant of motor cortical activity during stimulus presentation and in the absence of changes in peripheral motor events, in contrast to the commonly held view of the motor cortex as just an "upper motor neuron."     

细胞松弛素B对牛卵母细胞体外成熟过程中微管、微丝及染色体的影响

细胞松弛素B(Cytochalasin B,CB)是一种抑制微丝聚合的药物,能抑制微丝的组装从而阻止胞质分裂和极体排放,是研究细胞分裂器形成与变化的重要药物。在牛卵母细胞体外成熟培养过程中加入7.5 μg/mL的CB进行处理,分析CB对减数分裂过程中细胞骨架形态、染色体的排列与分离等方面的影响。结果显示,CB处理后卵母细胞第一极体的排放受到了抑制,染色体的排列和分离受到了影响,出现了同源染色体分离不完全或分离不均匀及分离后又聚在一起等异常情况,形成许多二倍体卵母细胞;纺锤体微管的形态发生了变化,出现了两个纺锤体、巨大纺锤体和多极纺锤体等异常结构;微丝的正常分布受到了影响,染色体周围没有或少有微丝分布,皮质下的微丝分布也变得少而不均匀;这说明微丝与微管在减数分裂过程中是协同作用的,CB通过影响微丝的动态变化,改变了纺锤体微管的形态结构,最终抑制了极体的排放。     

Distinguishing inchworm and hand-over-hand processive kinesin movement by neck rotation measurements

The motor enzyme kinesin makes hundreds of unidirectional 8-nanometer steps without detaching from or freely sliding along the microtubule on which it moves. We investigated the kinesin stepping mechanism by immobilizing a Drosophila kinesin derivative through the carboxyl-terminal end of the neck coiled-coil domain and measuring orientations of microtubules moved by single enzyme molecules at submicromolar adenosine triphosphate concentrations. The kinesin-mediated microtubule-surface linkage was sufficiently torsionally stiff (>/=2.0 +/- 0.9 x 10(-20) Newton meters per radian2) that stepping by the hypothesized symmetric hand-over-hand mechanism would produce 180 degree rotations of the microtubule relative to the immobilized kinesin neck. In fact, there were no rotations, a finding that is inconsistent with symmetric hand-over-hand movement. An alternative "inchworm" mechanism is consistent with our experimental results.     

一种地区分销系统的利润优化模型及其算法

研究由一个地区分销中心和多个不同的零售商组成的地区分销系统，假设顾客需求是随机的价格弹性需求．在基于价格折扣的联合决策供应链契约下。由地区分销中心提供一个价格折扣，以系统各成员追求利润最大化为目标，建立了地区分销系统的利润优化模型。并给出了一种基于遗传算法的求解策略．最后，通过算例说明该模型的有效性．     

Cytoplasmic dynein moves through uncoordinated stepping of the AAA+ ring domains

Cytoplasmic dynein is a homodimeric AAA+ motor that transports a multitude of cargos toward the microtubule minus end. How the two catalytic head domains interact and move relative to each other during processive movement is unclear. Here, we tracked the relative positions of both heads with nanometer precision and directly observed the heads moving independently along the microtubule. The heads remained widely separated, and their stepping behavior varied as a function of interhead separation. One active head was sufficient for processive movement, and an active head could drag an inactive partner head forward. Thus, dynein moves processively without interhead coordination, a mechanism fundamentally distinct from the hand-over-hand stepping of kinesin and myosin.     

似锥低颈吸虫的精子和精子形成的超微结构研究（复殖目：棘口科）

似锥低颈吸虫（Ｈｙｐｏｄｅｒａｅｕｍｃｏｎｏｉｄｅｕｍ）的成熟精子分为前段和主段。前段含两条鞭毛轴丝和一圈连续纵向排列的膜下微管；主段含两条不等长鞭毛轴丝、核、线粒体、糖原，背侧和腹侧的膜下微管。在精子形成过程中，精子细胞出现胞质突起，核与胞质突起部的质膜之间出现特殊分化区域（ＺＤ）。特殊分化区域的组成包括质膜下一排膜下微管，两个具有横纹小根的中心粒和１个中心粒间体（Ｉｎｔｅｒｃｅｎｔｒｉｏｌａｎｂｏｄｙ）。鞭毛从中心粒伸出，延伸，与中间不断伸长的胞质突起平行，核旋转，拉长，与细长的线粒体前后进入胞质突起，最后鞭毛从近端到远端（Ｐｒｏｘｉｍｉｔｏｄｉｓｔａｌｌｙ）与中间胞质突起融合，并从精子细胞群上脱落，形成成熟精子，遗留下精子细胞群的残体。残体（Ｒｅｓｉｄｕａｌｂｏｄｙ）上含弓形膜结构，线粒体，鞭毛轴丝残余以及电子致密颗粒，在一个残体上，至少有１０个以上的精子断端。     

The molecular architecture of axonemes revealed by cryoelectron tomography

Eukaryotic flagella and cilia are built on a 9 + 2 array of microtubules plus >250 accessory proteins, forming a biological machine called the axoneme. Here we describe the three-dimensional structure of rapidly frozen axonemes from Chlamydomonas and sea urchin sperm, using cryoelectron tomography and image processing to focus on the motor enzyme dynein. Our images suggest a model for the way dynein generates force to slide microtubules. They also reveal two dynein linkers that may provide "hard-wiring" to coordinate motor enzyme action, both circumferentially and along the axoneme. Periodic densities were also observed inside doublet microtubules; these may contribute to doublet stability.     

猪笼草叶笼滑移区各向异性超疏水润湿特性表征与机理分析

为获取各向异性超疏水表面研制的仿生原型,采用接触角测量仪测试表征了猪笼草(Nepenthes alata)叶笼滑移区的各向异性超疏水润湿特性,采用扫描电镜和三维形貌干涉仪观测滑移区形貌结构并提取三维特征信息,分析滑移区形貌结构对各向异性超疏水润湿特性的影响机理。结果表明:水滴在滑移区的接触角为(155.07±1.14)°,朝向叶笼底部和顶部的滚动角分别为(2.82±0.45)°和(5.40±0.31)°;滑移区覆盖着两端朝向叶笼内部弯曲且具有不对称凸面表层轮廓的微米级月骨体,以及形貌轮廓不规则且交错排列成致密孔洞结构的纳米级蜡质晶体,其中能够蓄留空气的蜡质晶体是产生超疏水润湿特性的重要因素;月骨体在不同方向的斜坡结构、悬崖结构致使水滴在滑移区滚动难易程度呈现差异,从而导致滚动角的显著不同与各向异性超疏水润湿现象的产生。     

甘蔗茎尖细胞有丝分裂过程中微管骨架的变化（英文）

In order to understand the microtubule change of monocotyls stem-tip during mitosis,the arrangement,transformation of microtubule array and its relation with chromosome movement during mitosis were studied with freezing microtome,indirect immunofluorescence,DAPI staining and fluorescence microscopy.The results showed that nucleolus was intact when the cortical microtubules formed;cortical microtubules were changed into phramoplast microtubule bands at mitosis prophase.When phramoplast microtubule came into being,nuclear membrane was ruptured and chromosome was arranged at the position of cell plate;subsequently,phramoplast microtubules were changed into phragmoplast microtubules,phramoplast microtubules were shortening and microtubules on the sides of cell plate were increasing gradually,during this course sister chromatid was separated by microtubules at cell plate and tract to the two poles,forming phragmoplast microtubules.Then the nucleolus of two daughter cells formed and separated in the end with the increase of cells numbers.Therefore,cell division orientation could be judged from the arrangement of cell microtubules in different periods in order to understand its growth status.