Human functional genetic studies are biased against the medically most relevant primate-specific genes

Background  

Many functional, structural and evolutionary features of human genes have been observed to correlate with expression breadth and/or gene age. Here, we systematically explore these correlations.     

Effects of varying Notch1 signal strength on embryogenesis and vasculogenesis in compound mutant heterozygotes

Background  

Identifying developmental processes regulated by Notch1 can be addressed in part by characterizing mice with graded levels of Notch1 signaling strength. Here we examine development in embryos expressing various combinations of Notch1 mutant alleles. Mice homozygous for the hypomorphic Notch1 ^12f allele, which removes the single O-fucose glycan in epidermal growth factor-like repeat 12 (EGF12) of the Notch1 ligand binding domain (lbd), exhibit reduced growth after weaning and defective T cell development. Mice homozygous for the inactive Notch1 ^lbd allele express Notch1 missing an ~20 kDa internal segment including the canonical Notch1 ligand binding domain, and die at embryonic day ~E9.5. The embryonic and vascular phenotypes of compound heterozygous Notch1 ^12f/lbd embryos were compared with Notch1 ^+/12f , Notch1 ^12f/12f , and Notch1 ^lbd/lbd embryos. Embryonic stem (ES) cells derived from these embryos were also examined in Notch signaling assays. While Notch1 signaling was stronger in Notch1 ^12f/lbd compound heterozygotes compared to Notch1 ^lbd/lbd embryos and ES cells, Notch1 signaling was even stronger in embryos carrying Notch1 ^12f and a null Notch1 allele.     

Plant Biodiversity in China: Richly Varied, Endangered, and in Need of Conservation

China is among the world's richest countries in terms of plant biodiversity. Besides the abundant flora, containing some 33,000 vascular plants (30,000 angiosperms, 250 gymnosperms, and 2600 pteridophytes), there is extraordinary ecosystem diversity, as well as a large pool of both wild and cultivated germplasms. China is also considered one of the main centers of origin and diversification for seed plants on Earth, and is especially profuse in phylogenetically primitive taxa and/or paleoendemics due to the refuge role glaciation played during the Quaternary period. The collision with the Indian subcontinent significantly enriched Chinese flora and led to the formation of many neoendemisms. However, flora distribution remains uneven, and some local floristic hotspots are found across China, such as Yunnan, Sichuan and Taiwan. Unfortunately, this biodiversity faces enormous threats, which have increased substantially over the last 50 years. The combined effects of habitat destruction and/or fragmentation, environmental contamination, over-exploitation of natural resources, and to a lesser extent, introduction of exotic species, have caused irreparable damage to China's plant biodiversity. Burgeoning economic and population growth have also contributed to this deterioration. It is believed that up to 5000 flora species are currently endangered in China, with some taxa having already become extinct. Although in recent years government authorities have made some efforts to preserve biodiversity, much work remains to be done. While China has established an extensive network of nature reserves and protected areas, encompassing more than 16% of the total land area, insufficient budgetary and staffing commitments are common limitations in their management structures. Ex-situ conservation is also deficient, primarily because the botanical gardens are not representative of several local floras, nor are they often of adequate size or representative of endangered species. The lack of effective and efficient environmental legislation and education are also problems that continue to accelerate the loss of plant biodiversity in China.     

The design and synthesis of a tricyclic single-nitrogen scaffold that serves as a 5-HT2C receptor agonist

5-HT2C agonists have shown efficacy in limiting food consumption and thus may serve as an important drug class in combating obesity. We describe the design and synthesis of a novel tricyclic single-nitrogen scaffold that was used to produce 5-HT2C agonists. SAR was developed around this chemotype and compounds were identified that were potent (Ki<15 nM) and selective relative to other 5-HT2 receptors. The most promising compound displayed a good pharmacokinetic profile in multiple animal species, and was efficacious in an acute feeding study in rats.     

Peptide deformylase is a potential target for anti-Helicobacter pylori drugs: reverse docking, enzymatic assay, and X-ray crystallography validation

Colonization of human stomach by the bacterium Helicobacter pylori is a major causative factor for gastrointestinal illnesses and gastric cancer. However, the discovery of anti-H. pylori agents is a difficult task due to lack of mature protein targets. Therefore, identifying new molecular targets for developing new drugs against H. pylori is obviously necessary. In this study, the in-house potential drug target database (PDTD, http://www.dddc.ac.cn/tarfisdock/) was searched by the reverse docking approach using an active natural product (compound 1) discovered by anti-H. pylori screening as a probe. Homology search revealed that, among the 15 candidates discovered by reverse docking, only diaminopimelate decarboxylase (DC) and peptide deformylase (PDF) have homologous proteins in the genome of H. pylori. Enzymatic assay demonstrated compound 1 and its derivative compound 2 are the potent inhibitors against H. pylori PDF (HpPDF) with IC50 values of 10.8 and 1.25 microM, respectively. X-ray crystal structures of HpPDF and the complexes of HpPDF with 1 and 2 were determined for the first time, indicating that these two inhibitors bind well with HpPDF binding pocket. All these results indicate that HpPDF is a potential target for screening new anti-H. pylori agents. In addition, compounds 1 and 2 were predicted to bind to HpPDF with relatively high selectivity, suggesting they can be used as leads for developing new anti-H. pylori agents. The results demonstrated that our strategy, reverse docking in conjunction with bioassay and structural biology, is effective and can be used as a complementary approach of functional genomics and chemical biology in target identification.     

Microbial ecology in the age of genomics and metagenomics: concepts, tools, and recent advances

Microbial ecology examines the diversity and activity of micro-organisms in Earth's biosphere. In the last 20 years, the application of genomics tools have revolutionized microbial ecological studies and drastically expanded our view on the previously underappreciated microbial world. This review first introduces the basic concepts in microbial ecology and the main genomics methods that have been used to examine natural microbial populations and communities. In the ensuing three specific sections, the applications of the genomics in microbial ecological research are highlighted. The first describes the widespread application of multilocus sequence typing and representational difference analysis in studying genetic variation within microbial species. Such investigations have identified that migration, horizontal gene transfer and recombination are common in natural microbial populations and that microbial strains can be highly variable in genome size and gene content. The second section highlights and summarizes the use of four specific genomics methods (phylogenetic analysis of ribosomal RNA, DNA-DNA re-association kinetics, metagenomics, and micro-arrays) in analysing the diversity and potential activity of microbial populations and communities from a variety of terrestrial and aquatic environments. Such analyses have identified many unexpected phylogenetic lineages in viruses, bacteria, archaea, and microbial eukaryotes. Functional analyses of environmental DNA also revealed highly prevalent, but previously unknown, metabolic processes in natural microbial communities. In the third section, the ecological implications of sequenced microbial genomes are briefly discussed. Comparative analyses of prokaryotic genomic sequences suggest the importance of ecology in determining microbial genome size and gene content. The significant variability in genome size and gene content among strains and species of prokaryotes indicate the highly fluid nature of prokaryotic genomes, a result consistent with those from multilocus sequence typing and representational difference analyses. The integration of various levels of ecological analyses coupled to the application and further development of high throughput technologies are accelerating the pace of discovery in microbial ecology.     

Seasonal change and habitat selection of shorebird community at the South Yangtze River Mouth and North Hangzhou Bay, China

Coastal regions are important habitats for migratory shorebirds. The aim of the study is to understand habitat use by migratory shorebirds and to develop a conservation strategy in the sustainable use of wetlands. From March 2004 to January 2005, we conducted a seasonal shorebirds census in ten coastal habitats along the South Yangtze River mouth and North Hangzhou Bay, simultaneously examining the relative seasonal abundance of shorebirds and their spatial distribution. A total of 25 species were identified, the dominant seasonal species were Great Knot (Calidris tenuirostris), Sharp-tailed Sandpiper (Calidris alpine) and Red-necked Stint (Calidris ruficollis) in spring; Kentish Plover (Charadrius alexandrinus), Common Greenshank(Tringa nebularia) and Lesser Sand Plover (Charadrius mongolus) in summer; Kentish Plover, Red-necked Stint and Common Greenshank in autumn; Dunlin(Calidris alpine), Kentish Plover and Marsh Sandpiper (Tringa stagnatilis) in winter. These species accounted for more than 85% of the total shorebirds. The numbers of shorebirds counted was highest in spring and then in autumn, winter and summer respectively. Among the four seasons, there were few significant differences in the number of bird species between the sites outside the seawall (intertidal mudflat) and the sites inside the seawall (artificial wetland), but the average density of shorebirds was obviously different. The habitat-selection analysis of the environmental factors (outside and inside the seawall) impacting on the shorebird community was made in the 10 study sites with Canonical Correspondence Analysis. The study results indicated that: (1) Outside the seawall, the widths of the total intertidal mudflat and bare mudflat were the key factors affecting the shorebirds; the proportion of bulrush (Scirpus×mariquete) covering and supertidal mudflat width had a positive correlation with the abundance of birds, while human disturbance and the proportion of both reed (Phragmites communis) and smooth cord-grass (Spartina alterniflora) covering in total surveyed areas had negative impacts on bird numbers; (2) Inside the seawall, the proportions of areas with shallow water and mudflats occupying the total surveyed area were key factors influencing the number of birds; the size of the bulrush area should have a positive impact on the appearance of shorebirds. Habitats with heavy human disturbance, dense reed and smooth cord-grass or a high water level were not conducive to be inhabited by shorebirds.     

Crystal structures of human NUDT5 reveal insights into the structural basis of the substrate specificity

Human NUDT5 (hNUDT5) is an ADP-ribose pyrophosphatase (ADPRase) belonging to the Nudix hydrolase superfamily. It presumably plays important roles in controlling the intracellular level of ADP-ribose (ADPR) to prevent non-enzymatic ADP-ribosylation by hydrolyzing ADPR to AMP and ribose 5'-phosphate. We report here the crystal structures of hNUDT5 in apo form, in complex with ADPR, and in complex with AMP with bound Mg2+. hNUDT5 forms a homodimer with substantial domain swapping and assumes a structure more similar to Escherichia coli ADPRase ORF209 than human ADPRase NUDT9. The adenine moiety of the substrates is specifically recognized by the enzyme via hydrogen-bonding interactions between N1 and N6 of the base and Glu47 of one subunit, and between N7 of the base and Arg51 of the other subunit, providing the molecular basis for the high selectivity of hNUDT5 for ADP-sugars over other sugar nucleotides. Structural comparisons with E. coli ADPRase ORF209 and ADPXase ORF186 indicate that the existence of an aromatic residue on loop L8 in ORF186 seems to be positively correlated with its enzymatic activity on APnA, whereas hNUDT5 and ORF209 contain no such residue and thus have low or no activities on APnA.     

Structural and molecular interactions of CCR5 inhibitors with CCR5

We have characterized the structural and molecular interactions of CC-chemokine receptor 5 (CCR5) with three CCR5 inhibitors active against R5 human immunodeficiency virus type 1 (HIV-1) including the potent in vitro and in vivo CCR5 inhibitor aplaviroc (AVC). The data obtained with saturation binding assays and structural analyses delineated the key interactions responsible for the binding of CCR5 inhibitors with CCR5 and illustrated that their binding site is located in a predominantly lipophilic pocket in the interface of extracellular loops and within the upper transmembrane (TM) domain of CCR5. Mutations in the CCR5 binding sites of AVC decreased gp120 binding to CCR5 and the susceptibility to HIV-1 infection, although mutations in TM4 and TM5 that also decreased gp120 binding and HIV-1 infectivity had less effects on the binding of CC-chemokines, suggesting that CCR5 inhibition targeting appropriate regions might render the inhibition highly HIV-1-specific while preserving the CC chemokine-CCR5 interactions. The present data delineating residue by residue interactions of CCR5 with CCR5 inhibitors should not only help design more potent and more HIV-1-specific CCR5 inhibitors, but also give new insights into the dynamics of CC-chemokine-CCR5 interactions and the mechanisms of CCR5 involvement in the process of cellular entry of HIV-1.