Chromatin-independent nuclear envelope assembly induced by Ran GTPase in Xenopus egg extracts

The nuclear envelope (NE) forms a controlled boundary between the cytoplasm and the nucleus of eukaryotic cells. To facilitate investigation of mechanisms controlling NE assembly, we developed a cell-free system made from Xenopus laevis eggs to study the process in the absence of chromatin. NEs incorporating nuclear pores were assembled around beads coated with the guanosine triphosphatase Ran, forming pseudo-nuclei that actively imported nuclear proteins. NE assembly required the cycling of guanine nucleotides on Ran and was promoted by RCC1, a nucleotide exchange factor recruited to beads by Ran-guanosine diphosphate (Ran-GDP). Thus, concentration of Ran-GDP followed by generation of Ran-GTP is sufficient to induce NE assembly.     

Enzymes without borders: mobilizing substrates, delivering products

Many cellular reactions involve both hydrophobic and hydrophilic molecules that reside within the chemically distinct environments defined by the phospholipid-based membranes and the aqueous lumens of cytoplasm and organelles. Enzymes performing this type of reaction are required to access a lipophilic substrate located in the membranes and to catalyze its reaction with a polar, water-soluble compound. Here, we explore the different binding strategies and chemical tricks that enzymes have developed to overcome this problem. These reactions can be catalyzed by integral membrane proteins that channel a hydrophilic molecule into their active site, as well as by water-soluble enzymes that are able to capture a lipophilic substrate from the phospholipid bilayer. Many chemical and biological aspects of this type of enzymology remain to be investigated and will require the integration of protein chemistry with membrane biology.     

The division of endosymbiotic organelles

Mitochondria and chloroplasts are essential eukaryotic organelles of endosymbiotic origin. Dynamic cellular machineries divide these organelles. The mechanisms by which mitochondria and chloroplasts divide were thought to be fundamentally different because chloroplasts use proteins derived from the ancestral prokaryotic cell division machinery, whereas mitochondria have largely evolved a division apparatus that lacks bacterial cell division components. Recent findings indicate, however, that both types of organelles universally require dynamin-related guanosine triphosphatases to divide. This mechanistic link provides fundamental insights into the molecular events driving the division, and possibly the evolution, of organelles in eukaryotes.     

荧光分光光度法测定大鼠不同脑区单胺类神经递质

介绍一种测定大鼠不同脑区单胺类神经递质的方法。采用荧光分光光度法同时测定大鼠不同脑区NE、DA和5-HT三种单胺类神经递质的含量。结果表明,荧光强度与浓度呈线性相关,NE、DA和5-HT的检测回收率分别为102.2%、95.4%和102.8%。该方法快速、灵敏、重复性较高,可为临床监测及科研提供参考。     

胞间连丝结构与组分的研究进展

胞间连丝是植物体内连接相邻细胞的一种跨细胞的细胞器,是细胞间物质运输和信息传递的通道,为相邻细胞间小分子、病毒颗粒及一些特殊大分子的运输提供途径。细胞间的共质通道就在质膜和其内部的压缩内质网间形成,压缩内质网又称连丝微管。由于胞间连丝结构的复杂性,使得对其结构组分的研究受到很大限制,但近几年随着电子显微镜、免疫标记、显微注射技术的广泛应用,已有证据表明肌动蛋白、肌球蛋白、中心蛋白和钙网蛋白等是胞间连丝的组分,这些蛋白之间相互作用调节胞间连丝的通透性,控制物质运输。     

Sequence homology between certain viral proteins and proteins related to encephalomyelitis and neuritis

Post-infectious or post-vaccinal demyelinating encephalomyelitis and neuritis may be due to immunological cross-reactions evoked by specific viral antigenic determinants (epitopes) that are homologous to regions in the target myelins of the central and peripheral nervous systems. Such homologies have been found by computer searches in which decapeptides in two human myelin proteins were compared with proteins of viruses known to infect humans. These viruses include measles, Epstein-Barr, influenza A and B, and others that cause upper respiratory infections. Several regions identified in myelin basic protein and P2 protein can be related to experimental allergic encephalomyelitis or neuritis in laboratory animals.     

Role of noradrenergic signaling by the nucleus tractus solitarius in mediating opiate reward

Norepinephrine (NE) is widely implicated in opiate withdrawal, but much less is known about its role in opiate-induced locomotion and reward. In mice lacking dopamine beta-hydroxylase (DBH), an enzyme critical for NE synthesis, we found that NE was necessary for morphine-induced conditioned place preference (CPP; a measure of reward) and locomotion. These deficits were rescued by systemic NE restoration. Viral restoration of DBH expression in the nucleus tractus solitarius, but not in the locus coeruleus, restored CPP for morphine. Morphine-induced locomotion was partially restored by DBH expression in either brain region. These data suggest that NE signaling by the nucleus tractus solitarius is necessary for morphine reward.     

减蛋综合征病毒NE4株全基因组序列测定与分析

根据减蛋综合征病毒(EDSV-76)AV-127株的全基因序列(序列号BK000404),用Premier5.0软件设计22对引物,利用PCR方法对EDSV-76病毒NE4株的全基因组进行了分段扩增、克隆和序列测定,并用DNAStar分析软件对各片段的测序结果进行拼接,将该序列与GenBank中已登录的相应序列进行同源性分析,并用六邻体蛋白构建进化树.结果表明:NE4株基因组序列全长33214bp,GC含量为43%,与国际标准株AV-127相比,核苷酸序列同源性为99.6%.碱基的插入或缺失主要在非编码区,在100K蛋白编码区距羧基端1/10处插入了1个碱基C,其ORF编码696个氨基酸,而AV-127株100K蛋白编码709个氨基酸,预计其发挥功能的基团在N端.蛋白同源性分析表明,EDSVNE4株主要蛋白与羊腺病毒OAV(Ovine adenovirusD)、牛腺病毒BAV(Bovine adenovi-rusD)和蛇腺病毒SAV(Snake adenovirus)有较高的同源性,而与禽腺病毒Ⅰ群(CELO)的同源性较低,说明NE4株与禽腺病毒Ⅰ群亲缘关系较远,进化树分析也证实了此特性.     

AT-hook蛋白的研究进展

AT-hook是一类新的DNA结合蛋白基序,与其他功能已知的DNA结合基序不同,AT-hook基序具有以精氨酸-甘氨酸-精氨酸-脯氨酸（RGRP）四个残基为中心的特征结构。AT-hook蛋白与DNA的特异结合是通过AT-hook基序的氨基酸残基与双链DNA小沟中富含AT碱基的区域相互作用完成的。AT-hook基序广泛存在于不同物种的DNA结合蛋白中,AT-hook蛋白在染色质结构组装和对目标基因转录活性的调控中起着重要的作用,进而影响生物的生长发育。     

Global identification of human transcribed sequences with genome tiling arrays

Elucidating the transcribed regions of the genome constitutes a fundamental aspect of human biology, yet this remains an outstanding problem. To comprehensively identify coding sequences, we constructed a series of high-density oligonucleotide tiling arrays representing sense and antisense strands of the entire nonrepetitive sequence of the human genome. Transcribed sequences were located across the genome via hybridization to complementary DNA samples, reverse-transcribed from polyadenylated RNA obtained from human liver tissue. In addition to identifying many known and predicted genes, we found 10,595 transcribed sequences not detected by other methods. A large fraction of these are located in intergenic regions distal from previously annotated genes and exhibit significant homology to other mammalian proteins.     

Axonopathy and transport deficits early in the pathogenesis of Alzheimer's disease

We identified axonal defects in mouse models of Alzheimer's disease that preceded known disease-related pathology by more than a year; we observed similar axonal defects in the early stages of Alzheimer's disease in humans. Axonal defects consisted of swellings that accumulated abnormal amounts of microtubule-associated and molecular motor proteins, organelles, and vesicles. Impairing axonal transport by reducing the dosage of a kinesin molecular motor protein enhanced the frequency of axonal defects and increased amyloid-beta peptide levels and amyloid deposition. Reductions in microtubule-dependent transport may stimulate proteolytic processing of beta-amyloid precursor protein, resulting in the development of senile plaques and Alzheimer's disease.     

Axonal transport: each major rate component reflects the movement of distinct macromolecular complexes

The proteins of the three major rate components of axonal transport in guinea pig retinal ganglion cells were analyzed by one- and two-dimensional gel electrophoresis. Each rate component consisted of a different set of proteins that remained associated with each other during transport. This suggests that each rate component represents a distinct macromolecular complex and that these complexes may be definable organelles such as microtubules, microfilaments, and smooth endoplasmic reticulum. Thus, the transport of radiolabeled proteins in the axon reflects the movement of complete subcellular rather than the movement of individual proteins.     

大学生英语需求分析——以右江民族医学院护理专业英语本科为例

为了解护理专业(英语方向)学生英语学习需求,探讨适合英语教学和学习的思路和方法。抽取右江民族医学院2008、2009级护英本学生作为本研究的对象,开展英语学习需求调查问卷和访谈。结果表明:学生对英语学习认同度高,对自身英语水平比较不满意,希望教师能够改进英语教学方法和教材,提高其英语实际应用能力。     

气候变化对东北沼泽湿地潜在分布的影响

东北地区是我国沼泽湿地分布最广泛的地区。为研究沼泽湿地对气候变化的响应,选取了对沼泽湿地分布可能存在影响的26个环境因子,利用最大熵(Maximum Entropy, MaxEnt)模型模拟了沼泽湿地基准气候条件下的潜在分布,并预测了气候变化情景下2011-2040 年、2041-2070 年和2071-2100 年3个研究阶段东北沼泽湿地潜在分布。研究结果表明:最大熵模型预测精度较高(平均AUC(Aera Under Curve)为(0.826±0.005))。基准气候条件下东北沼泽潜在分布区主要为大小兴安岭和三江平原地区。随着时间的推进,东北地区沼泽湿地原有潜在分布面积明显减少,而新增潜在分布面积较少,总面积呈现急剧减少趋势。至2071-2100年,原有沼泽湿地潜在分布面积将减少99.80%,新增潜在分布面积仅2.48%,总潜在分布面积减少97.32%。空间分布上,东北沼泽湿地潜在分布呈现由东向西迁移,南北向中心收缩的趋势。研究结果可为东北地区沼泽湿地保护政策的制定提供参考。     

Binding of ARF and beta-COP to Golgi membranes: possible regulation by a trimeric G protein

The binding of cytosolic coat proteins to organelles may regulate membrane structure and traffic. Evidence is presented that a small guanosine triphosphate (GTP)-binding protein, the adenosine diphosphate ribosylation factor (ARF), reversibly associates with the Golgi apparatus in an energy, GTP, and fungal metabolite brefeldin A (BFA)-sensitive manner similar to, but distinguishable from, the 110-kilodalton cytosolic coat protein beta-COP. Addition of beta gamma subunits of G proteins inhibited the association of both ARF and beta-COP with Golgi membranes that occurred upon incubation with guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S). Thus, heterotrimeric G proteins may function to regulate the assembly of coat proteins onto the Golgi membrane.     

Hydrophobic organization of membrane proteins

Membrane-exposed residues are more hydrophobic than buried interior residues in the transmembrane regions of the photosynthetic reaction center from Rhodobacter sphaeroides. This hydrophobic organization is opposite to that of water-soluble proteins. The relative polarities of interior and surface residues of membrane and water soluble proteins are not simply reversed, however. The hydrophobicities of interior residues of both membrane and water-soluble proteins are comparable, whereas the bilayer-exposed residues of membrane proteins are more hydrophobic than the interior residues, and the aqueous-exposed residues of water-soluble proteins are more hydrophilic than the interior residues. A method of sequence analysis is described, based on the periodicity of residue replacement in homologous sequences, that extends conclusions derived from the known atomic structure of the reaction center to the more extensive database of putative transmembrane helical sequences.     

Conversion of a PI-anchored protein to an integral membrane protein by a single amino acid mutation

Qa-2, a cell-surface glycoprotein anchored by phosphatidylinositol (PI), is structurally related to the class I transplantation antigens H-2 K, D, and L, which are integral membrane glycoproteins. The predicted transmembrane segment of Qa-2 differs from those of H-2 K, D, and L by the presence of an aspartate in place of a valine at position 295. A single base change that replaced this aspartate with valine resulted in cell-surface Qa-2 molecules that were insensitive to hydrolysis by a PI-specific phospholipase C and more resistant to papain cleavage, properties shared by H-2D. Cells expressing Asp----Val mutant Qa-2 proteins were still able to attach a PI anchor to endogenous proteins such as Thy-1 and J11D. It therefore appears that this single amino acid change converts Qa-2 from a PI-linked form into an integral membrane protein.     

Uncoupling translocation from translation: implications for transport of proteins across membranes

The segregation of secretory proteins into the cisternae of the endoplasmic reticulum (ER) is normally tightly coupled to their synthesis. This feature distinguishes their biogenesis from that of proteins targeted to many other organelles. In the examples presented, translocation across the ER membrane is dissociated from translation. Transport, which is normally cotranslational, may proceed in the absence of chain elongation. Moreover, translocation across the ER membrane does not proceed spontaneously since, even in the absence of protein synthesis, energy substrates are required for translocation. These conclusions have been extended to the cotranslational integration of newly synthesized transmembrane proteins.