Retrograde axonal transport of endogenous proteins in sciatic nerve demonstrated by covalent labeling in vivo

Extracellularly applied N-succinimidyl [2,3-3H]propionate was used in vivo to covalently label intra-axonal proteins in the rat sciatic nerve. This technique permitted a unique view of axonal transport of proteins independent of biosynthesis. The proteins detected in slow anterograde transport (1 to 2 millimeters per day) correspond to cytoskeletal proteins described in previous papers. The slowly retrogradely transported component (3 to 6 millimeters per day) was composed primarily of a single protein with a molecular weight of 68,000.     

P1 of strawberry vein banding virus,a multilocalized protein,functions as a movement protein and interacts with the coat protein

Although the complete nucleotide sequence of strawberry vein banding virus (SVBV) has been determined and bioinformatic analysis has revealed that the SVBV genome could encode seven proteins, the precise function of each protein is unclear. This study provided evidence that the P1 protein of SVBV (SVBV-P1) possesses the following features. Bioinformatic and subcellular localization analyses showed that SVBV-P1 is localized in the cytoplasm and cell walls of epidermal cells in Nicotiana benthamiana, and it forms inclusion bodies associated with microtubules and the endoplasmic reticulum. Dilution experiments demonstrated that SVBV-P1 could move from the original agro-infiltrated cells to adjacent cells in N. benthamiana leaves. Further trans-complementation experiments demonstrated that SVBV-P1 could facilitate the intercellular movement of a movement-deficient potato virus X mutant in N. benthamiana leaves. Finally, yeast two-hybrid and bimolecular fluorescence complementation assays revealed that SVBV-P1 could interact with the SVBV coat protein, which is a major component of Caulimovirus virions. Results of the electrophoretic mobility shift assay indicated that SVBV-P1 lacks DNA-binding capability. In summary, the results suggest that SVBV-P1 is probably a movement protein of SVBV, providing new insights into the function of movement proteins of the Caulimovirus genus.     

Rapid axonal transport of sulfated mucopolysaccharide proteins

When sulfur-35-labeled sodium sulfate is injected intraocularly in the goldfish, labeled sulfated mucopolysaccharides rapidly appear in the contra-lateral optic tectum of the brain, demonstrating the axonal flow of sulfated mucopolysaccharides. The transport rate is the same as that observed for proteins labeled after intraocular injection of tritiated proline. Treatment of the sulfur-35-labeled material with precipitants and enzymes reveals the presence of substances with properties similar to those of heparan sulfate (the major component) and chondroitin sulfate. Dermatan sulfate was not detected.     

杜仲ABC转运蛋白基因家族成员鉴定与分析

ABC转运蛋白(ATP—binding cassette transpoter)是一类庞大而古老的跨膜运输蛋白家族,在生物体内参与多种物质的转运积累、有害物质解毒、气孔调节、植物防御等生理活动。杜仲(Eucommia ulmoides Oliv)作为重要的中药材,其药用成分主要为次生代谢产物,次生代谢物转运与积累过程需要ABC转运蛋白的参与。利用生物信息学手段对杜仲ABC转运蛋白基因家族进行鉴定,并分析该家族蛋白质性质和结构、跨膜结构、亚细胞定位、系统进化关系。研究表明,EuABC家族生物信息学预测有76个成员,含有1~7个保守基序;编码蛋白多为稳定蛋白,主要分布于细胞质膜上,二级结构以α-螺旋和无规卷曲为主要构成元件;进化树分析表明,杜仲ABC转运蛋白家族可分为8个亚家族(A~G;I),每组成员数量分别为3、19、14、1、1、1、29、8。研究结果可为进一步研究杜仲次生代谢物质转运与积累奠定基础,也为其他植物ABC转运蛋白家族的研究提供了参考依据。     

Transport of protein by goldfish optic nerve fibers

After tritiated leucine was injected into the eye of goldfish, radio-active protein synthesized by the ganglion cell bodies moved down the optic axons at an average rate of 0.4 mm per day. Radioautograms of the optic tectum in which these axons end show that, as early as 24 hours after the injection, before the radioactivity in the tectal layer containing the optic axons had risen above background level, the layer containing the axon terminals was already heavily labeled. The radioactivity in the terminals reached a maximum about 48 hours after the injection and remained approximately constant for at least 23 days thereafter, whereas the radioactivity in the fiber layer increased significantly during the same interval, as the slowly moving protein component entered it. Thus there appears to be a special mechanism for rapid transport of protein from the cell body to the synaptic terminals, as well as a slower movement of protein down the axon.     

影响我国气旋活动的几个重要因素

气旋是大气中常见的重要天气系统,它的发生、发展和移动对于各个地区、各纬度之间的热量交换、水汽输送和广大地区的天气变化有着很大的影响。了解它们的发展、演变和移动规律,对天气分析和预报是很重要的。     

Direct visualization of organelle movement along actin filaments dissociated from characean algae

A system has been developed in which organelle transport can be studied without the influence of an organized cellular cytoplasm. Binding and continuous unidirectional movement of organelles along isolated cellular transport cables were directly visualized by video light microscopy after the dissociation of the cytoplasm of characean algae cells in a Ca2+-free buffer containing adenosine triphosphate. Individual organelles had more than one attachment site and moved at mean rates of 11.2 or 62.1 micrometers per second along multiple parallel pathways on each cable. Electron microscopy of these cables after direct freezing demonstrated that they consist of compact bundles of actin filaments. Under these conditions, characteristics of organelle movement should reflect directly the underlying molecular processes of binding and force generation.     

脂筏在病原微生物侵入细胞中的作用

研究表明,许多不同类型和种属的病原微生物利用脂筏结构介导进入细胞,完成转运和复制。对脂筏在病原微生物侵入细胞中的作用进行了研究,有助于探索新的抗病原微生物进入细胞的途径。     

利用酵母双杂交系统筛选玉米ZmPRR73的互作蛋白

为了阐明ZmPRR73基因的生物学功能,构建诱饵表达载体pGBKT7-ZmPRR73,利用酵母双杂交技术,从长日照光照环境诱导的热带玉米自交系的cDNA文库中筛选与ZmPRR73互作的蛋白。结果显示：构建的诱饵载体pGBKT7-ZmPRR73对酵母菌株无毒性,且对报告基因无自激活活性,共鉴定出12个与ZmPRR73互作的候选蛋白。生物信息学分析表明,这些候选互作蛋白的功能涉及植物的转录调控、离子跨膜转运的调节、信号转导、电子传递链等多个方面,推测ZmPRR73蛋白与以上蛋白互作参与多个信号转导和代谢途径,研究结果补充和完善了ZmPRR73蛋白参与的调控途径,为进一步研究生物钟核心元件ZmPRR73的分子功能提供了新的分子证据。     

Role of a highly conserved bacterial protein in outer membrane protein assembly

After transport across the cytoplasmic membrane, bacterial outer membrane proteins are assembled into the outer membrane. Meningococcal Omp85 is a highly conserved protein in Gram-negative bacteria, and its homolog Toc75 is a component of the chloroplast protein-import machinery. Omp85 appeared to be essential for viability, and unassembled forms of various outer membrane proteins accumulated upon Omp85 depletion. Immunofluorescence microscopy revealed decreased surface exposure of outer membrane proteins, which was particularly apparent at the cell-division planes. Thus, Omp85 is likely to play a role in outer membrane protein assembly.     

4UM-640型振动式马铃薯挖掘机的设计与试验

为适应甘肃省马铃薯种植地块小、坡地多的现状,提高马铃薯机械化收获水平,通过对各关键作业部件进行理论分析与设计选型,研制出了一种携有偏心摇臂结构振动挖掘装置,挖掘深度可调节且与手扶拖拉机相配套的4UM-640型振动式马铃薯挖掘机。田间试验表明：该挖掘机收获作业时掘薯、输运顺畅,减阻碎土、土薯分离效果明显,伤薯率为3.6%,损失率为4.1%,明薯率为97.1%,达到了马铃薯机械化收获的相关技术要求。     

植物质膜内在水通道蛋白PIPs的分子生物学研究进展

水是植物细胞的重要组成成分,植物水通道蛋白是细胞间和细胞内水分快速运输的主要通道,作为植物水通道蛋白的一个亚类,质膜内在蛋白PIPs定位于原生质膜,为典型的高水分选择性通道蛋白。主要介绍了PIPs的结构特征、生物学功能及其调控机制。高等植物PIPs存在2个高度保守的区域:GGGANXXXXGY和TGI/TNPARSL/FGAAI/VI/VFWF/YN。PIPs分为PIP1和PIP2亚类,PIP1比PIP2具较长的N－端和较短的C－端,并且具有各自的保守氨基酸。通过转基因和非洲爪蟾卵Xenopus oocytes母细胞异源表达研究表明PIPs不仅是水和二氧化碳、甘油等中性小分子选择性通道蛋白,同时还具有许多生理功能,是一类多功能蛋白。蛋白翻译后修饰、异聚化、pH值、二价阳离子等都能调控PIPs的运输功能。表1参51     

Methylation increases sodium transport into A6 apical membrane vesicles: possible mode of aldosterone action

When isolated apical membrane vesicles prepared from cultured A6 epithelia were incubated in vitro with the methyl donor S-adenosylmethionine, the control rate of amiloride-inhibitable sodium transport was doubled. The methylation inhibitors 3-deazaadenosine and S-adenosyl homocysteine returned the S-adenosyl-methionine-stimulated sodium transport to control levels. Neither these agents nor adenosine affected sodium transport into control vesicles. In vesicles incubated with S-adenosyl-[3H-methyl]methionine, both membrane phospholipids and proteins were labeled, and this labeling was inhibited by deazaadenosine. In vesicles prepared from A6 cells treated with aldosterone, sodium transport was twice the control value and S-adenosylmethionine did not cause any further stimulation of transport. In those vesicles, both lipid and protein methylation were increased. These results suggest that methylation, which increases the rate of amiloride-sensitive sodium transport is involved in the action of aldosterone at the apical membrane level in epithelia.     

Acropetal movement of auxin: dependence on temperature

The amounts of radioactive indole-3-acetic acid that moved, both basipetally and acropetally, through short sections of bean epicotyl were measured at 15 degrees and 25 degrees C in the presence and absence of 2,3,5-triiodobenzoic acid. Low temperature and this second compound each inhibited acropetal efflux of indole 3-acetic acid more than they did the influx. Results are interpreted as supporting a metabolically dependent component of the acropetal movement of auxin in bean epicotyls.     

Microtubule gelation-contraction: essential components and relation to slow axonal transport

Preparations of microtubule proteins isolated by assembly and disassembly undergo gelation-contraction after addition of adenosine triphosphate (ATP). A particulate fraction from these preparations that is required, along with purified tubulin, to produce ATP-dependent microtubule gelation-contraction in vitro has been isolated. The particulates exhibited microtubule-stimulated adenosine triphosphatase activity and moved slowly (about 1 micrometer per minute) along microtubule walls in the presence of ATP. The particulates contained tubulin, neurofilament, and spectrin polypeptides. The composition, solubility, and motility of the particulates are consistent with those of slow component a of axonal transport.     

原茬地大豆垄上双行开沟器结构参数优化

针对大豆垄上双行免耕播种作业过程中开沟器入土困难、开沟器堵塞、种沟构建质量差等问题,设计原茬地大豆垄上双行开沟器结构参数优化试验。通过分析工作过程中土壤颗粒在导流板表面运动规律,确定影响土壤颗粒运动规律关键结构与作业参数。采用四因素三水平正交试验方法,以分土角、运土角、导流角为试验因素,开沟深度变异系数和横向粒距变异系数为评价指标。优化结果为在作业速度8 km·h-1条件下,分土角65°,运土角30°,导流角45°时,开沟深度变异系数0.091、横向粒距变异系数0.035,作业过程无堵塞现象。将结构参数优化后双行开沟器与分体式双圆盘开沟器作比较试验,结果表明,开沟深度合格率提高11%,种子粒距横向分布合格率提高32%,开沟深度变异系数降低43.5%,横向粒距变异系数降低66.6%。研究结果为原茬地作物双行免耕播种机具推广应用提供技术支撑。     

Bacterial motility: membrane topology of the Escherichia coli MotB protein

The MotB protein of Escherichia coli is an essential component of the force generators that couple proton movement across the cytoplasmic membrane to rotation of the flagellar motors. The membrane topology of MotB was examined to explore the possibility that it might form a proton channel. MotB--alkaline phosphatase fusion proteins were constructed to identify likely periplasmic domains of the MotB molecule. Fusions distal to a putative membrane-spanning segment near the amino terminus of MotB exhibited alkaline phosphatase activity, indicating that an extensive carboxyl-terminal portion of MotB may be located on the periplasmic side of the membrane. Protease treatment of MotB in spheroplasts confirmed this view. The simple transmembrane organization of MotB is difficult to reconcile with a role as a proton conductor.     

Rates of surficial rock creep on hillslopes in Western colorado

The average rate of downslope movement of rock fragments on shale hillslopes is directly proportional to the sine of the slope angle or that component of the gravitational force which acts parallel to the hillslope. The rates of surficial rock creep range from a few millimeters per year on a 3degree slope to almost 70 millimeters per year on a 40-degree slope, but these rates vary with natural variations in soil characteristics and microclimate, as well as with accidental disturbances.     

安徽省高标准生态农田建设管理问题研究

研究了温度对甜瓜穴盘苗根际水分含量、可吸收水分比例及幼苗根系总根长、根系表面积、根系活力、气孔导度等生理指标的影响。结果表明,高、低温处理均会对甜瓜幼苗水分运移产生影响:高温处理6d后基质内水分散失剧烈,自由水比例迅速下降至4.46%,增加了甜瓜幼苗根系对水分吸收利用的难度;高、低温处理致使甜瓜幼苗根长、根系表面积等指标显著下降,根系脯氨酸含量上升、根系活力下降,从而影响了幼苗水分吸收;高、低温处理下叶片气孔导度、蒸腾速率显著低于常温处理,水分在植物体内的运移受到了一定的限制。     

Kinesin and dynein move a peroxisome in vivo: a tug-of-war or coordinated movement?

We used fluorescence imaging with one nanometer accuracy (FIONA) to analyze organelle movement by conventional kinesin and cytoplasmic dynein in a cell. We located a green fluorescence protein (GFP)-tagged peroxisome in cultured Drosophila S2 cells to within 1.5 nanometers in 1.1 milliseconds, a 400-fold improvement in temporal resolution, sufficient to determine the average step size to be approximately 8 nanometers for both dynein and kinesin. Furthermore, we found that dynein and kinesin do not work against each other in vivo during peroxisome transport. Rather, multiple kinesins or multiple dyneins work together, producing up to 10 times the in vitro speed.