Visualizing transient events in amino-terminal autoprocessing of HIV-1 protease

HIV-1 protease processes the Gag and Gag-Pol polyproteins into mature structural and functional proteins, including itself, and is therefore indispensable for viral maturation. The mature protease is active only as a dimer with each subunit contributing catalytic residues. The full-length transframe region protease precursor appears to be monomeric yet undergoes maturation via intramolecular cleavage of a putative precursor dimer, concomitant with the appearance of mature-like catalytic activity. How such intramolecular cleavage can occur when the amino and carboxy termini of the mature protease are part of an intersubunit beta-sheet located distal from the active site is unclear. Here we visualize the early events in N-terminal autoprocessing using an inactive mini-precursor with a four-residue N-terminal extension that mimics the transframe region protease precursor. Using paramagnetic relaxation enhancement, a technique that is exquisitely sensitive to the presence of minor species, we show that the mini-precursor forms highly transient, lowly populated (3-5%) dimeric encounter complexes that involve the mature dimer interface but occupy a wide range of subunit orientations relative to the mature dimer. Furthermore, the occupancy of the mature dimer configuration constitutes a very small fraction of the self-associated species (accounting for the very low enzymatic activity of the protease precursor), and the N-terminal extension makes transient intra- and intersubunit contacts with the substrate binding site and is therefore available for autocleavage when the correct dimer orientation is sampled within the encounter complex ensemble.     

Reversal of pathological pain through specific spinal GABAA receptor subtypes

Inflammatory diseases and neuropathic insults are frequently accompanied by severe and debilitating pain, which can become chronic and often unresponsive to conventional analgesic treatment. A loss of synaptic inhibition in the spinal dorsal horn is considered to contribute significantly to this pain pathology. Facilitation of spinal gamma-aminobutyric acid (GABA)ergic neurotransmission through modulation of GABA(A) receptors should be able to compensate for this loss. With the use of GABA(A)-receptor point-mutated knock-in mice in which specific GABA(A) receptor subtypes have been selectively rendered insensitive to benzodiazepine-site ligands, we show here that pronounced analgesia can be achieved by specifically targeting spinal GABA(A) receptors containing the alpha2 and/or alpha3 subunits. We show that their selective activation by the non-sedative ('alpha1-sparing') benzodiazepine-site ligand L-838,417 (ref. 13) is highly effective against inflammatory and neuropathic pain yet devoid of unwanted sedation, motor impairment and tolerance development. L-838,417 not only diminished the nociceptive input to the brain but also reduced the activity of brain areas related to the associative-emotional components of pain, as shown by functional magnetic resonance imaging in rats. These results provide a rational basis for the development of subtype-selective GABAergic drugs for the treatment of chronic pain, which is often refractory to classical analgesics.     

Genotype, haplotype and copy-number variation in worldwide human populations

Genome-wide patterns of variation across individuals provide a powerful source of data for uncovering the history of migration, range expansion, and adaptation of the human species. However, high-resolution surveys of variation in genotype, haplotype and copy number have generally focused on a small number of population groups. Here we report the analysis of high-quality genotypes at 525,910 single-nucleotide polymorphisms (SNPs) and 396 copy-number-variable loci in a worldwide sample of 29 populations. Analysis of SNP genotypes yields strongly supported fine-scale inferences about population structure. Increasing linkage disequilibrium is observed with increasing geographic distance from Africa, as expected under a serial founder effect for the out-of-Africa spread of human populations. New approaches for haplotype analysis produce inferences about population structure that complement results based on unphased SNPs. Despite a difference from SNPs in the frequency spectrum of the copy-number variants (CNVs) detected--including a comparatively large number of CNVs in previously unexamined populations from Oceania and the Americas--the global distribution of CNVs largely accords with population structure analyses for SNP data sets of similar size. Our results produce new inferences about inter-population variation, support the utility of CNVs in human population-genetic research, and serve as a genomic resource for human-genetic studies in diverse worldwide populations.     

环境工程实验教学方法的探索与实践

环境工程实验课程是环境工程及相关专业学生理论联系实际的桥梁之一,在专业人才培养中地位重要.根据该课程开设的目的和要求,为高质量地培养学生的实践动手能力、探索和创新思想以及严谨务实的科学作风,对该课程的教学手段和方法已开展了7年的相关探索与实践,取得了较好的教学效果.     

基于漏洞知识库的8031单片机系统软件漏洞检测算法

针对基于8031单片机系统软件的安全问题,对各权威漏洞数据库进行了分析研究,采用一种基于ECV规则的攻击分析方法从攻击事件中提取漏洞知识,根据漏洞种类及特征将漏洞从代码安全的角度分类,设计了三层结构的漏洞知识库,并根据漏洞知识库的设计提出了一种基于知识的漏洞检测算法,用于检测8031单片机系统漏洞。基于上述方法设计并实现了软件安全性逆向分析系统,对8031单片机系统进行漏洞检测。实验结果表明,基于该漏洞知识库的漏洞检测算法可以对目标程序正确进行漏洞检测,有利于降低软件代码漏洞量,并在一定程度上降低成本和资源消耗。      

杭州湾地区生态-气候监测预警系统及其应用

基于3S技术、中尺度模式及动态植被模式建立了生态-气候监测预警系统,其中包括4个子系统:土地覆盖动态监测系统、城市热岛动态监测系统、城市气候监测系统和植被NPP模拟与监测系统.利用土地覆盖监测系统可以精确地获取各种土地覆盖类型,并分析土地利用覆盖变化;利用土地覆盖监测系统结果、Landsat-TM红外影像及常规气象资料构建城市热岛监测系统,分析该地区城市热岛效应的时空变化:利用土地覆盖分类结果,对改变下垫面进行敏感性试验,模拟城市发展对城市气候的影响,构建城市气候监测系统:利用LPJ模拟杭州湾地区植被NPP及对气候变化的响应,构建植被NPP模拟与监测系统.生态预警系统可以实时监测土地覆盖、城市热岛、城市气候、植被NPP变化.对保护耕地资源、改善生态环境、提高城市人居环境、合理利用气候资源、保持农林业的可持续发展有着重要的指导和现实意义.     

明帝国何以错失海洋时代

历史学家公认,近代世界始于地理大发现。哥伦布（1492年）、达·伽马（1497年）、麦哲伦（1519年）完成的“地理大发现”,翻开了人类史新的一页,正是从这个时期开始,才有了全球史,而只有在有了全球史的情况下,才可能有全球战略。     

房地产工程合同策划

房地产工程项目的合同策划,是指在法律许可范围内,根据房地产公司总的目标要求,通过对合同条款做合理、完善的策划,使项目在建设活动的时间、资金使用关系中选择最佳结合点,保证项目获得满意的经济效益和社会效益.     

Mechanisms of sperm-egg interactions emerging from gene-manipulated animals

Untangling the molecular nature of sperm-egg interactions is fundamental if we are to understand fertilization. These phenomena have been studied for many years using biochemical approaches such as antibodies and ligands that interact with sperm or with eggs and their vestments. However, when homologous genetic recombination techniques were applied, most of the phenotypic factors of the gene-manipulated animals believed “essential” for fertilization were found to be dispensable. Of course, all biological systems contain redundancies and compensatory mechanisms, but as a whole the old model of fertilization clearly requires significant modification. In this review, we use the results of gene manipulation experiments in animals to propose the basis for a new vision. Received 26 January 2007; received after revision 7 March 2007; accepted 17 April 2007     

Mutations in amphiphysin 2 (BIN1) disrupt interaction with dynamin 2 and cause autosomal recessive centronuclear myopathy

Centronuclear myopathies are characterized by muscle weakness and abnormal centralization of nuclei in muscle fibers not secondary to regeneration. The severe neonatal X-linked form (myotubular myopathy) is due to mutations in the phosphoinositide phosphatase myotubularin (MTM1), whereas mutations in dynamin 2 (DNM2) have been found in some autosomal dominant cases. By direct sequencing of functional candidate genes, we identified homozygous mutations in amphiphysin 2 (BIN1) in three families with autosomal recessive inheritance. Two missense mutations affecting the BAR (Bin1/amphiphysin/RVS167) domain disrupt its membrane tubulation properties in transfected cells, and a partial truncation of the C-terminal SH3 domain abrogates the interaction with DNM2 and its recruitment to the membrane tubules. Our results suggest that mutations in BIN1 cause centronuclear myopathy by interfering with remodeling of T tubules and/or endocytic membranes, and that the functional interaction between BIN1 and DNM2 is necessary for normal muscle function and positioning of nuclei.