70K heat shock related proteins stimulate protein translocation into microsomes

A yeast cytosol is shown to contain two distinct activities that stimulate protein translocation across microsomal membranes. One activity was purified. It consists of two constitutively expressed 70K heat shock related proteins that increase the rate of translocation. Possible mechanisms of action of these proteins are discussed.     

Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots

The roots of most higher plants form arbuscular mycorrhiza, an ancient, phosphate-acquiring symbiosis with fungi, whereas only four related plant orders are able to engage in the evolutionary younger nitrogen-fixing root-nodule symbiosis with bacteria. Plant symbioses with bacteria and fungi require a set of common signal transduction components that redirect root cell development. Here we present two highly homologous genes from Lotus japonicus, CASTOR and POLLUX, that are indispensable for microbial admission into plant cells and act upstream of intracellular calcium spiking, one of the earliest plant responses to symbiotic stimulation. Surprisingly, both twin proteins are localized in the plastids of root cells, indicating a previously unrecognized role of this ancient endosymbiont in controlling intracellular symbioses that evolved more recently.     

全基因组预测樟疫霉的候选效应分子

为了更好地研究樟疫霉的致病机制及防治方法,笔者基于病原菌效应分子具有的典型特征,利用Signal P、Prot Comp、TMHMM、big-PI Fungal Predictor和Target P等生物信息学预测程序对樟疫霉中328 457条蛋白质序列进行候选效应分子找寻,发现该菌含有3 439个小分子分泌蛋白,同时,对上述蛋白进行冗余性、半胱氨酸数量以及信号肽长度等性质进行分析。结果表明:上述分泌蛋白中存在较多的冗余性蛋白,所占比例为50%以上,并以含有1~10个半胱氨酸、17~26个氨基酸信号肽的分泌蛋白居多。另外,利用卵菌中效应分子所具有的保守基序RXLR,对拥有唯一氨基酸序列的1 549个分泌蛋白进行基序找寻,明确樟疫霉中存在160个候选效应分子。通过上述生物信息学分析方法可实现樟疫霉候选效应分子的预测。     

Assembly of yeast Sec proteins involved in translocation into the endoplasmic reticulum into a membrane-bound multisubunit complex

Secretory-protein translocation into the endoplasmic reticulum (ER) is thought to be catalysed by integral membrane proteins. Genetic selections uncovered three Saccharomyces cerevisiae genes (SEC61, SEC62 and SEC63), mutations in which block import of precursor proteins into the ER lumen in vivo and in vitro. The DNA sequences of SEC62 and SEC63 predict multispanning membrane proteins, and biochemical characterization of the SEC62 protein (Sec62) confirms that it is an integral ER membrane protein. Here we show that Sec61, Sec62 and Sec63 are assembled with two additional proteins into a multisubunit membrane-associated complex. These results confirm previous predictions, based upon genetic interactions between the SEC genes, that Sec61, Sec62 and Sec63 act together to facilitate protein translocation into the ER.     

植物细胞钙靶蛋白的研究进展

简述了植物细胞的钙离子调控过程，对钙调蛋白、钙调磷酸酶B类蛋白和钙离子依赖型蛋白质激酶3类钙靶蛋白的最新研究进展进行了综述。     

埃博拉病毒入侵宿主细胞的分子机制

 埃博拉病毒是一类能够感染并引起人和灵长类动物发生埃博拉出血热的烈性囊膜病毒。发现近40年中,埃博拉病毒给人类生命带来了极大威胁。然而,目前人们对于埃博拉病毒的了解非常有限,尤其是病毒与其宿主细胞受体的结合机制和膜融合机制相关信息的缺失,使得针对埃博拉病毒的特效药物的设计和研发工作阻碍重重。本文综述了埃博拉病毒分类、形态、病毒蛋白和病毒生命周期,着重介绍了高福院士团队在埃博拉病毒入侵宿主细胞的分子机制研究中的成果。通过结构学手段解析了埃博拉病毒激活态囊膜糖蛋白GPcl与宿主细胞受体NPC1分子的复合物结构,从原子水平上阐明了埃博拉病毒与宿主细胞相互识别的机制,并在结构基础上对病毒的膜融合促发机制做出推测,提出以埃博拉病毒为代表的新的（第5种）囊膜病毒膜融合激发机制,为抗埃博拉病毒药物和疫苗的设计提供了结构基础。     

A substrate-specific inhibitor of protein translocation into the endoplasmic reticulum

The segregation of secretory and membrane proteins to the mammalian endoplasmic reticulum is mediated by remarkably diverse signal sequences that have little or no homology with each other. Despite such sequence diversity, these signals are all recognized and interpreted by a highly conserved protein-conducting channel composed of the Sec61 complex. Signal recognition by Sec61 is essential for productive insertion of the nascent polypeptide into the translocation site, channel gating and initiation of transport. Although subtle differences in these steps can be detected between different substrates, it is not known whether they can be exploited to modulate protein translocation selectively. Here we describe cotransin, a small molecule that inhibits protein translocation into the endoplasmic reticulum. Cotransin acts in a signal-sequence-discriminatory manner to prevent the stable insertion of select nascent chains into the Sec61 translocation channel. Thus, the range of substrates accommodated by the channel can be specifically and reversibly modulated by a cell-permeable small molecule that alters the interaction between signal sequences and the Sec61 complex.     

Protein delivery into eukaryotic cells by type III secretion machines

Bacteria that have sustained long-standing close associations with eukaryotic hosts have evolved specific adaptations to survive and replicate in this environment. Perhaps one of the most remarkable of those adaptations is the type III secretion system (T3SS)--a bacterial organelle that has specifically evolved to deliver bacterial proteins into eukaryotic cells. Although originally identified in a handful of pathogenic bacteria, T3SSs are encoded by a large number of bacterial species that are symbiotic or pathogenic for humans, other animals including insects or nematodes, and plants. The study of these systems is leading to unique insights into not only organelle assembly and protein secretion but also mechanisms of symbiosis and pathogenesis.     

无机纳米颗粒在植物转化中的应用

随着纳米生物技术的发展,基于纳米材料构建基因载体的植物转基因技术,逐渐成为一类具有划时代意义的创新性高效植物转基因技术。笔者综述了羟基磷灰石、硅、碳纳米管、量子点、磁性纳米颗粒等无机纳米颗粒在植物转化中的应用,并比较了这些纳米载体的优势及存在的问题。分析认为,不同纳米材料对受体植物细胞的影响、纳米材料及其构建的载体入胞机制等基础理论问题迫切需要进一步阐明,入胞途径的细胞生物学和生理生化过程需要进一步实证,开发可定向输送目的基因到特定细胞或细胞器的安全高效新载体、目的基因的高效释放和功能激发等,将是未来一段时间内纳米植物生物技术研究的主要方向。     

一种快速准确区分III型、IV型分泌效应蛋白的计算方法

通过Ⅲ型、Ⅳ型、Ⅵ型分泌系统,革兰氏阴性菌可将效应蛋白直接注入宿主体内,并导致宿主感染各种疾病.由于Ⅲ型、Ⅳ型分泌效应蛋白均属非经典分泌蛋白,且它们可能具有相似的序列模体或进化保守性,故两者之间难于区分.基于支持向量机和伪位置特异性得分矩阵,本文提出了一种可快速准确识别革兰氏阴性菌Ⅲ型、Ⅳ型效应蛋白的计算方法.测试集实验结果表明,本方法对Ⅲ型、Ⅳ型效应蛋白具有较好的分类效果,可作为辅助工具用于分泌效应蛋白的进一步研究.     

The structural basis for activation of plant immunity by bacterial effector protein AvrPto

Pathogenic microbes use effectors to enhance susceptibility in host plants. However, plants have evolved a sophisticated immune system to detect these effectors using cognate disease resistance proteins, a recognition that is highly specific, often elicits rapid and localized cell death, known as a hypersensitive response, and thus potentially limits pathogen growth. Despite numerous genetic and biochemical studies on the interactions between pathogen effector proteins and plant resistance proteins, the structural bases for such interactions remain elusive. The direct interaction between the tomato protein kinase Pto and the Pseudomonas syringae effector protein AvrPto is known to trigger disease resistance and programmed cell death through the nucleotide-binding site/leucine-rich repeat (NBS-LRR) class of disease resistance protein Prf. Here we present the crystal structure of an AvrPto-Pto complex. Contrary to the widely held hypothesis that AvrPto activates Pto kinase activity, our structural and biochemical analyses demonstrated that AvrPto is an inhibitor of Pto kinase in vitro. The AvrPto-Pto interaction is mediated by the phosphorylation-stabilized P+1 loop and a second loop in Pto, both of which negatively regulate the Prf-mediated defences in the absence of AvrPto in tomato plants. Together, our results show that AvrPto derepresses host defences by interacting with the two defence-inhibition loops of Pto.     

A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides

Depletion of a subset of 70K stress proteins in yeast mutants shows that they are involved in the post-translational import of precursor polypeptides into both mitochondria and the lumen of the endoplasmic reticulum. The identification of such a basic function may explain the remarkable evolutionary conservation of the gene family encoding these proteins.     

Bioorganic synthesis of lipid-modified proteins for the study of signal transduction

Biological membranes define the boundaries of the cellular compartments in higher eukaryotes and are active in many processes such as signal transduction and vesicular transport. Although post-translational lipid modification of numerous proteins in signal transduction is crucial for biological function, analysis of protein-protein interactions has mainly focused on recombinant proteins in solution under defined in vitro conditions. Here we present a new strategy for the synthesis of such lipid-modified proteins. It involves the bacterial expression of a carboxy-terminally truncated non-lipidated protein, the chemical synthesis of differently lipidated peptides representing the C terminus of the proteins, and their covalent coupling. Our technique is demonstrated using Ras constructs, which exhibit properties very similar to fully processed Ras, but can be produced in high yields and are open for selective modifications. These constructs are operative in biophysical and cellular assay systems, showing specific recognition of effectors by Ras lipoproteins inserted into the membrane surface of biosensors and transforming activity of oncogenic variants after microinjection into cultured cells.     

Requirement for hsp70 in the mitochondrial matrix for translocation and folding of precursor proteins

By analysis of a temperature-sensitive yeast mutant, a heat-shock protein in the matrix of mitochondria, mitochondrial hsp70 (Ssc1p), is found to be involved both in translocation of nuclear-encoded precursor proteins across the mitochondrial membranes and in (re)folding of imported proteins in the matrix.     

Single-molecule fluorescence imaging of membrane-bound proteins for studies of cell signal transduction

Fluorescence imaging of single molecules is becoming a powerful tool to examine biological processes at the molecular level.Using total internal reflection fluorescence microscopy (TIRFM),it has been possible to study the dynamic behavior of single molecules on living cell membranes.Herein,we briefly review the application of TIRFM-based single-molecule imaging in studies of membrane receptors involved in signal transduction.Furthermore,we discuss several examples of our own research on growth factor receptors,including TGF-β receptors,HER2,and EGFR,and speculate possible applications of this technique to investigate other cellular events occurring on or near the plasma-membrane.     

Role of acetylcholine on plant root-shoot signal transduction

The role of accetylcholine (ACh) on plant root-shoot communication was investigated using the root-split system of Vicia faba L.In the experiments,slight osmotic stress caused the decrease of ACh content in root tips and the xylem sap transported up per time unit from root tip to the shool when the water potential of the shoot was kept unchanged .It also caused the decrease of ACh content in the abaxial epidermis,The decrease was highly correlative to the changes of transpiration rate,suggesting that the decrase of ACh content probably functions as a signal to regulate stomatal behavior,The effect of osmotic stress might be mainly through the inhibition of the ACh synthesis in root tip ;thus further influences the ACh content in root tip ,xylem sap and abaxial epidermis and resulting in the changes of stomata behavior,These results provide new evdence that plants transduce positive and negative signals among roots and shoot to coordinate stomatal behavior and adapt to variable environments.     

Retinal bipolar cells——A model for the study of neuronal signal integration

Being the functional basis of neural networks, neuronal signal integration has been currently becoming a hot point of neuroscience research. Active and passive membrane properties of retinal bipolar cells, temporal and spatial distribution of synaptic inputs to these cells are described, and modulation of signal integration characteristics of these cells under different retinal adaptation states is analyzed. In consideration of the functional and morphological properties of retinal bipolar cells, it is suggested that they may be an appropriate model for the study of neuronal signal integration.