A coupled model of fast axonal transport of organelles and slow axonal transport of tau protein

We have developed a model that accounts for the effect of a non-uniform distribution of tau protein along the axon length on fast axonal transport of intracellular organelles. The tau distribution is simulated by using a slow axonal transport model; the numerically predicted tau distributions along the axon length were validated by comparing them with experimentally measured tau distributions reported in the literature. We then developed a fast axonal transport model for organelles that accounts for the reduction of kinesin attachment rate to microtubules by tau. We investigated organelle transport for two situations: (1) a uniform tau distribution and (2) a non-uniform tau distribution predicted by the slow axonal transport model. We found that non-uniform tau distributions observed in healthy axons (an increase in tau concentration towards the axon tip) result in a significant enhancement of organelle transport towards the synapse compared with the uniform tau distribution with the same average amount of tau. This suggests that tau may play the role of being an enhancer of organelle transport.     

细胞的“新细胞器”——迁移体

迁移体（migrasome）是俞立教授于2015年报道的新细胞器。迁移体是细胞迁移过程中尾部产生的收缩丝的尖端或交叉点产生出的膜性细胞器。细胞产生迁移体的过程称为迁移性胞吐（migracytosis）,介导细胞内物质的释放和细胞间远距离通讯,在斑马鱼胚胎发育及器官形成中具有重要作用。本篇综述总结了目前有关迁移体的研究进展,包括早期迁移体的发现过程,TSPAN4和胆固醇形成的宏结构域,整合素(integrin)与细胞外基质的相互作用以及特异性是迁移体发生的核心分子机制、迁移体研究的第一个活体动物模型以及迁移体具有和潜在的生理意义、血清中迁移体的研究。本篇综述还归纳了当前建立的迁移体研究方法和工具,包括迁移体纯化的方法、迁移体的鉴定方法、迁移体的分子标志物、迁移体的染料标记方法和抑制迁移体发生的小分子抑制剂等相关研究进展,为迁移体领域的研究奠定工具基础和树立标准。本综述还对迁移体这个新兴领域中的重要问题和研究方向进行展望,期待更多其他领域的科学家投入迁移体领域的研究中。     

Interaction of Molecular Motors

This review is devoted to the problem of regulation of intracellular organelle transport along microtubules. Properties of the system of microtubules as transport tracks are considered. Some motor proteins (kinesins and dyneins) carrying their “cargoes” in opposite directions are characterized. Special attention is paid to the analysis of transport of the organelles that can change the direction of movement. Possible molecular mechanisms coordinating the functioning of opposite motor proteins are considered.__________Translated from Molekulyarnaya Biologiya, Vol. 39, No. 4, 2005, pp. 709–718.Original Russian Text Copyright © 2005 by Gyoeva.I dedicate this review to the memory of Andrei Nikolaevich Belozerskii. I had no opportunity to become his disciple because he had already passed away when I came to the Department of Plant Biochemistry, Biological Faculty, Moscow State University. Nevertheless, my education is indebted to the Department that he headed for a long time and whose life was charged with the charm of his personality.     

研究报告：改造颈部铰链区重塑驱动蛋白3的持续运动能力

目的 驱动蛋白3的颈部螺旋与其后的蛋白质序列相互关联,形成一个延长的颈部,其中包含一个特征铰链结构。该特征性铰链在不同的驱动蛋白中表现出多样性,在驱动蛋白KIF13B中,这个铰链仅由一个脯氨酸残基组成,而在驱动蛋白KIF1A中,则由一个长的柔性无规卷曲构成。然而,这个颈部铰链在控制驱动蛋白持续运动方面的功能仍不明确。方法 本文对KIF13B和KIF1A的颈部铰链区的氨基酸残基进行突变改造,并通过单分子运动实验研究铰链区突变对驱动蛋白运动行为的影响。结果 在KIF13B中,在铰链区——脯氨酸前后插入柔性残基对其运动的速度以及持续性都有不同程度的影响,而去除该脯氨酸则可以同时提高运动速度和持续性。在KIF1A中,删除整个柔性颈部铰链仅仅增强了其运动的持续性。同时,把驱动蛋白1的颈部铰链区用改造后的驱动蛋白3的颈部铰链区进行替换,同样能够提高驱动蛋白1的持续运动能力。结论 驱动蛋白3颈部铰链控制其持续运动能力,适当改造可以调整马达的运动行为,这为重塑驱动蛋白马达的持续运动提供了一种新策略。     

Two distinct regions in a yeast myosin-V tail domain are required for the movement of different cargoes

The Saccharomyces cerevisiae myosin-V, Myo2p, is essential for polarized growth, most likely through transport of secretory vesicles to the developing bud. Myo2p is also required for vacuole movement, a process not essential for growth. The globular region of the myosin-V COOH-terminal tail domain is proposed to bind cargo. Through random mutagenesis of this globular tail, we isolated six new single point mutants defective in vacuole inheritance, but not polarized growth. These point mutations cluster to four amino acids in an 11-amino acid span, suggesting that this region is important for vacuole movement. In addition, through characterization of myo2-DeltaAflII, a deletion of amino acids 1,459-1,491, we identified a second region of the globular tail specifically required for polarized growth. Whereas this mutant does not support growth, it complements the vacuole inheritance defect in myo2-2 (G1248D) cells. Moreover, overexpression of the myo2-DeltaAflII globular tail interferes with vacuole movement, but not polarized growth. These data indicate that this second region is dispensable for vacuole movement. The identification of these distinct subdomains in the cargo-binding domain suggests how myosin-Vs can move multiple cargoes. Moreover, these studies suggest that the vacuole receptor for Myo2p differs from the receptor for the essential cargo.     

Recent developments in chloroplast protein transport

Most proteins located in chloroplasts are encoded by nuclear genes, synthesized in the cytoplasm, and transported into the organelle. The study of protein uptake by chloroplasts has greatly expanded over the past few years. The increased activity in this field is due, in part, to the application of recombinant DNA methodology to the analysis of protein translocation. Added interest has also been gained by the realization that the transport mechanisms that mediate protein uptake by chloroplasts, mitochondria and the endoplasmic reticulum display certain characteristics in common. These include amino terminal sequences that target proteins to particular organelles, a transport process that is mechanistically independent from the events of translation, and an ATP-requiring transport step that is thought to involve partial unfolding of the protein to be translocated. In this review we examine recent studies on the binding of precursors to the chloroplast surface, the energy-dependent uptake of proteins into the stroma, and the targeting of proteins to the thylakoid lumen. These aspects of protein transport into chloroplasts are discussed in the context of recent studies on protein uptake by mitochondria.Abbrevlations CAT chloramphenicol acetyl transferase - CCCP carbonylcyanide m-chlorophenylhydrazone - DHFR dihydrofolate reductase - EPSP 5-enol-pyruvylshikimate-3-phosphate - ER endoplasmic reticulum - LHCP light harvesting chlorophyll a/b apoprotein - NPT neomycin phosphotransferase - oATP adenosine-2,3-dialdehyde-5-triphosphate - P-inorganic phosphate Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - SSU small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase - SRP signal recognition particle     

Selective protein transport: Identity of the solubilized phosvitin receptor from chicken oocytes

By two independent methods, the solubilized receptor for phosvitin (PV) has a subunit MW of 116K. Affinity chromatography, showed that only 2 of the more than 25 proteins present in the total detergent solubilized oocyte membrane extract were retained on a PV–agarose column. These proteins of MW of 116K and 100K could be eluted from PV–agarose with free PV. By gel exclusion chromatography, the receptor-¹²⁵I-PV complexes elute in the void volume of a Biogel A-1.5 column. When these void fractions were assayed by SDS-PAGE only a single protein of MW of 116K was observed in addition to ¹²⁵I-PV.     

Human chromokinesin KIF4A functions in chromosome condensation and segregation

Accurate chromosome alignment at metaphase and subsequent segregation of condensed chromosomes is a complex process involving elaborate and only partially characterized molecular machinery. Although several spindle associated molecular motors have been shown to be essential for mitotic function, only a few chromosome arm--associated motors have been described. Here, we show that human chromokinesin human HKIF4A (HKIF4A) is an essential chromosome-associated molecular motor involved in faithful chromosome segregation. HKIF4A localizes in the nucleoplasm during interphase and on condensed chromosome arms during mitosis. It accumulates in the mid-zone from late anaphase and localizes to the cytokinetic ring during cytokinesis. RNA interference--mediated depletion of HKIF4A in human cells results in defective prometaphase organization, chromosome mis-alignment at metaphase, spindle defects, and chromosome mis-segregation. HKIF4A interacts with the condensin I and II complexes and HKIF4A depletion results in chromosome hypercondensation, suggesting that HKIF4A is required for maintaining normal chromosome architecture. Our results provide functional evidence that human KIF4A is a novel component of the chromosome condensation and segregation machinery functioning in multiple steps of mitotic division.     

An RNA‐binding atypical tropomyosin recruits kinesin‐1 dynamically to oskar mRNPs

Localization and local translation of oskar mRNA at the posterior pole of the Drosophila oocyte directs abdominal patterning and germline formation in the embryo. The process requires recruitment and precise regulation of motor proteins to form transport‐competent mRNPs. We show that the posterior‐targeting kinesin‐1 is loaded upon nuclear export of oskar mRNPs, prior to their dynein‐dependent transport from the nurse cells into the oocyte. We demonstrate that kinesin‐1 recruitment requires the DmTropomyosin1‐I/C isoform, an atypical RNA‐binding tropomyosin that binds directly to dimerizing oskar 3′UTRs. Finally, we show that a small but dynamically changing subset of oskar mRNPs gets loaded with inactive kinesin‐1 and that the motor is activated during mid‐oogenesis by the functionalized spliced oskar RNA localization element. This inefficient, dynamic recruitment of Khc decoupled from cargo‐dependent motor activation constitutes an optimized, coordinated mechanism of mRNP transport, by minimizing interference with other cargo‐transport processes and between the cargo‐associated dynein and kinesin‐1.     

Plasmodesmata transport of GFP alone or fused to potato virus X TGBp1 is diffusion driven

Summary. Plasmodesmata (Pd) provide a pathway for exchanging various macromolecules between neighboring plant cells. Researchers routinely characterize the mobility of the green-fluorescent protein (GFP) and GFP fusions through Pd by calculating the proportion of sites in bombarded leaves which show fluorescence in multiple cell clusters (% movement). Here, the Arrhenius equation was used to describe the temperature dependence of GFP and GFP-TGBp1 (potato virus X triple gene block protein1) movement, using % movement values, and to calculate the activation energy for protein transport. The resulting low activation energy indicates GFP and GFP-TGBp1 movement are diffusion driven. Furthermore, GFP movement is inversely proportional to the leaf surface area of expanding leaves. The increase in leaf area results mainly from cell expansion during the sink–source transition. The increasing cell size results in lower Pd density, which decreases the probability that a GFP attains an open Pd by diffusion. The decline in GFP movement as leaf area expands indicates that, in addition to GFP diffusion through Pd, attaining an open Pd by undirected diffusion might be limiting for Pd transport. In summary, this report provides a new quantitative method for studying Pd conductivity. Correspondence: Jeanmarie Verchot Lubicz, Department of Entomology and Plant Pathology, 127 Noble Research Center, Oklahoma State University, Stillwater, OK 74078, U.S.A.     

Differences between migrasome,a ‘new organelle’, and exosome

The migrasome is a new organelle discovered by Professor Yu Li in 2015. When cells migrate, the membranous organelles that appear at the end of the retraction fibres are migrasomes. With the migration of cells, the retraction fibres which connect migrasomes and cells finally break. The migrasomes detach from the cell and are released into the extracellular space or directly absorbed by the recipient cell. The cytoplasmic contents are first transported to the migrasome and then released from the cell through the migrasome. This release mechanism, which depends on cell migration, is named ‘migracytosis’. The main components of the migrasome are extracellular vesicles after they leave the cell, which are easy to remind people of the current hot topic of exosomes. Exosomes are extracellular vesicles wrapped by the lipid bimolecular layer. With extensive research, exosomes have solved many disease problems. This review summarizes the differences between migrasomes and exosomes in size, composition, property and function, extraction method and regulation mechanism for generation and release. At the same time, it also prospects for the current hotspot of migrasomes, hoping to provide literature support for further research on the generation and release mechanism of migrasomes and their clinical application in the future.     

强壮果蝇(D.virilis)多线染色体的DNA·RNA内源杂交分子原位免疫酶标观察

应用兔抗polydT·polyrA的特异性抗体,在果蝇D.virilis多线染色体上用免疫酶标的方法检测到DNA·RNA内源杂交分子。多线染色体制片必须先进行变性复性处理(即内源分子杂交技术)才能得到明显的酶标显色带。大致可分辨到363条带,遍及染色体的各个区域。用DNA·RNA杂交分子为特异性底物的RNA酶H处理后再经变性复性处理后的多线染色体,免疫酶标的阳性带几乎全部消失,因此证明经内源分子杂交技术处理后用免疫化学方法检测到的阳性带确是由DNA·RNA内源杂交分子所构成的。     

Separating speed and ability to displace roadblocks during DNA translocation by FtsK

FtsK translocates dsDNA directionally at >5 kb/s, even under strong forces. In vivo, the action of FtsK at the bacterial division septum is required to complete the final stages of chromosome unlinking and segregation. Despite the availability of translocase structures, the mechanism by which ATP hydrolysis is coupled to DNA translocation is not understood. Here, we use covalently linked translocase subunits to gain insight into the DNA translocation mechanism. Covalent trimers of wild‐type subunits dimerized efficiently to form hexamers with high translocation activity and an ability to activate XerCD‐dif chromosome unlinking. Covalent trimers with a catalytic mutation in the central subunit formed hexamers with two mutated subunits that had robust ATPase activity. They showed wild‐type translocation velocity in single‐molecule experiments, activated translocation‐dependent chromosome unlinking, but had an impaired ability to displace either a triplex oligonucleotide, or streptavidin linked to biotin‐DNA, during translocation along DNA. This separation of translocation velocity and ability to displace roadblocks is more consistent with a sequential escort mechanism than stochastic, hand‐off, or concerted mechanisms.     

水霉Saprolegnia菌丝内细胞颗粒运动的特征

水霉菌丝内细胞颗粒的运动有布朗运动和跳跃运动两种形式,跳跃运动的速度在０．０９至４μｍ／ｓ之间。细胞颗粒运动的速度时制改变．不是匀速运动。在同一根菌丝内细胞颗粒运动的速度与颗粒大小无明显相关性;在不同的菌丝内细胞颗粒运动的速度差异明显。细胞颗粒有共同的运动轨道,运动轨道弯曲,并与细胞长轴基本平行。在同一运动轨道上,不同细胞颗粒的运动速度不同。     

Comparison of Efficency of Translation Between a Deformable Swimmer Versus a Rigid Body in a Bounded Fluid Domain: Consequences for Subcellular Transport

In this paper, we compare the translation efficiencies of a deformable circle that swims by means of low amplitude periodic tangential surface waves versus a rigid circle, moving in a bounded fluid domain. The swimmer is found to be much more efficient than the rigid body. We believe that this result gives some support to the active hypothesis of subcellular transport, where it is supposed that the organelle can generate by itself a propulsive flux, (by changes of form or metabolic activities) instead of just being carried by the motion of an external agent, like a molecular motor.