原钙粘蛋白基因簇调控区域中成簇的CTCF结合位点分析

哺乳动物中原钙粘蛋白(Protocadherin, Pcdh)基因簇包含50多个串联排列的基因,这些基因形成3个紧密相连的基因簇(Pcdhα、Pcdhβ和Pcdhγ),所编码的原钙粘蛋白质群在神经元多样性(Neuronal diversity)和单细胞特异性(Single cell identity)以及神经突触信号转导中发挥重要作用。前期的工作已证实转录因子CTCF(CCCTC-binding factor)与CTCF结合位点(CTCF-binding site, CBS)的方向性结合能够决定增强子和启动子环化的方向以及其远距离交互作用的特异性,并进一步在Pcdh基因座(Locus)形成两个(Pcdhα和Pcdhγ)染色质拓扑结构域(CTCF/cohesin- mediated chromatin domain, CCD),而且染色质拓扑结构域对于控制基因表达调控至关重要。本文通过生物信息学方法对比人类和小鼠序列,发现Pcdhβγ染色质拓扑结构域调控区域中的DNase I超敏位点(DNase I hypersensitive sites, HSs)较为保守。染色质免疫沉淀及大规模测序实验(Chromatin immunoprecipitation and massive parallel sequencing, ChIP-Seq)揭示CBS位点在Pcdhβγ调控区域中成簇分布并且具有相同的方向。凝胶电泳迁移实验(Electrophoresis mobility shift assay, EMSA)确定Pcdhβγ调控区域内具体的42 bp CBS位点并且发现一个CTCF峰包含两个CBS位点。在全基因组范围内,运用计算生物学方法分析CTCF和增强子、启动子等调控元件的关系,发现CBS位点在调控元件附近有较多分布,推测CTCF通过介导增强子和启动子的特异性交互作用,在细胞核三维基因组内形成活性转录枢纽调控基因精准表达。     

The biomass and aboveground net primary productivity of <Emphasis Type="Italic">Schima superba-Castanopsis carlesii</Emphasis> forests in east China

The biomass and productivity of Schima superba-Castanopsis carlesii forests in Tiantong, Zhejiang Province, were determined using overlapping quadrants and stem analyses. The total community biomass was (225.3±30.1) t hm⁻², of which the aboveground parts accounted for 72.0% and the underground parts accounted for 28.0%. About 87.2% of biomass existed in the tree layer. The resprouting biomass was small, of which over 95.0% occurred in the shrub layer. The productivity of the aboveground parts of the community was (386.8±98.9) g m⁻²a⁻¹, in which more than 96.0% was present at the tree level. The trunk’s contribution to productivity was the greatest, while that of leaves was the smallest. In China, the community biomass of subtropical evergreen broadleaved forests differs significantly with the age of the forest. The community biomass of the 52-year-old S. superba-C. carlesii forests in this study was lower than the average biomass of subtropical evergreen broadleaved forests in China, and was lower than the biomass of other subtropical evergreen broadleaved forests elsewhere in the world. Moreover, its productivity was lower than the model estimate, indicating that without disturbance, this community has great developmental potential in terms of community biomass and productivity.     

Role of extracellular polymeric substance (EPS) in toxicity response of soil bacteria Bacillus sp. S3 to multiple heavy metals

Heavy metal resistant bacteria are of great interest because of their potential use in bioremediation. Understanding the survival and adaptive strategies of these bacteria under heavy metal stress is important for better utilization of these bacteria in remediation. The objective of this study was to investigate the role of bacterial extracellular polymeric substance (EPS) in detoxifying against different heavy metals in Bacillus sp. S3, a new hyper antimony-oxidizing bacterium previously isolated from contaminated mine soils. The results showed that Bacillus sp. S3 is a multi-metal resistant bacterial strain, especially to Sb(III), Cu(II) and Cr(VI). Toxic Cd(II), Cr(VI) and Cu(II) could stimulate the secretion of EPS in Bacillus sp. S3, significantly enhancing the adsorption and detoxification capacity of heavy metals. Both Fourier transform infrared spectroscopy (FTIR) and three-dimensional excitation–emission matrix (3D-EEM) analysis further confirmed that proteins were the main compounds of EPS for metal binding. In contrast, the EPS production was not induced under Sb(III) stress. Furthermore, the TEM–EDX micrograph showed that Bacillus sp. S3 strain preferentially transported the Sb(III) to the inside of the cell rather than adsorbed it on the extracellular surface, indicating intracellular detoxification rather than extracellular EPS precipitation played an important role in microbial resistance towards Sb(III). Together, our study suggests that the toxicity response of EPS to heavy metals is associated with difference in EPS properties, metal types and corresponding environmental conditions, which is likely to contribute to microbial-mediated remediation.

     

肿瘤坏死因子－α（TNF－α）对常见呼吸道病毒的抑制作用

用人重组肿瘤坏死因子－α（Ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ－α,ＴＮＦ－α）和人天然α干扰素（Ｉｎｔｅｒｆｅｒｏｎ－α－,ＩＦＮ－α）在人胚胎肺纤维母细胞（ＨＥＦ）和Ｈｅｐ－２细胞系上对常见呼吸道病毒所致细胞病变抑制进行比较观察。病毒包括不同型别的腺病毒５株,单疱病毒Ⅰ型（ＨＳＶ－Ｉ）１株,鼻病毒１株,仙台病毒１株,ＶＳＶ１株。结果提示ＴＮＦ－α和ＩＦＮ－α均具有广谱抗病毒活性。ＴＮＦ－α的抑毒作用能被ＴＮＦ－α申抗和ＩＦＮ－β单抗完全去除,被ＩＦＮ－α单抗部分去除ＴＮＦ－α的抗病毒效应。ＴＮＦ－－α中和试验的结果提示：ＴＮＦ抗病毒活性仍为ＩＦＮ－β诱生所介导。     

Bose-Einstein condensation in biological systems

A theoretical model is presented for describing a previously untreated effect of viscosity on the apparent decomposition rate of enzyme-ligand complexes.Since the translational diffusion is hindered by the viscosity, its increased value results in an enlarged portion of ligands which can be rebound by the enzyme immediately after the dissociation of the complex.The model accounts for the experimentally observed decrease in maximal velocity of enzymic reactions at high viscosity. At the same time, it serves as a tool to obtain new information about the energetic processes of enzyme action.     

Abamectin resistance in Drosophila is related to increased expression of P-glycoprotein via the dEGFR and dAkt pathways

Many insects have evolved resistance to abamectin but the mechanisms involved in this resistance have not been well characterized. P-glycoprotein (P-gp), an ATP-dependent drug-efflux pump transmembrane protein, may be involved in abamectin resistance. We investigated the role of P-gp in abamectin (ABM) resistance in Drosophila using an ABM-resistant strain developed in the laboratory. A toxicity assay, Western blotting analysis and a vanadate-sensitive ATPase activity assay all demonstrated the existence of a direct relationship between P-gp expression and ABM resistance in these flies. Our observations indicate that P-gp levels in flies' heads were higher than in their thorax and abdomen, and that both P-gp levels and LC₅₀ values were higher in resistant than in susceptible and P-gp-deficient strains. In addition, P-gp levels in the blood–brain barrier (BBB) of resistant flies were higher than in susceptible and P-gp-deficient flies, which is further evidence that a high level of P-gp in the BBB is related to ABM resistance. Furthermore, we found greater expression of Drosophila EGFR (dEGFR) in the resistant strain than in the susceptible strain, and that the level of Drosophila Akt (dAkt) was much higher in resistant than in susceptible flies, whereas that in P-gp-deficient flies was very low. Compared to susceptible flies, P-gp levels in the resistant strain were markedly suppressed by the dEGFR and dAkt inhibitors lapatinib and wortmannin. These results suggest that the increased P-gp in resistant flies was regulated by the dEGFR and dAkt pathways and that increased expression of P-gp is an important component of ABM resistance in insects.     

Histidine ammonia-lyase from Streptomyces griseus.

Histidine ammonia-lyase (histidase; HutH) has been purified to homogeneity from Streptomyces griseus and the N-terminal amino acid (aa) sequence used to clone the histidase-encoding structural gene, hutH. The purified enzyme shows typical saturation kinetics and is inhibited competitively by D-histidine and histidinol phosphate. High concentrations of K.cyanide inactivate HutH unless the enzyme is protected by the substrate or histidinol phosphate. On the basis of the nucleotide sequence, the hutH structural gene would encode a protein of 53 kDa with an N terminus identical to that determined for the purified enzyme. Immediately upstream from hutH is a region that strongly resembles a class of Streptomyces promoters active during vegetative growth; however, there is no obvious ribosome-binding site adjacent to the hutH translation start codon. The deduced aa sequence of an upstream partial open reading frame shows no similarity with other proteins, including HutP of Bacillus subtilis and HutU of Pseudomonas putida. Promoter-probe analysis indicates that promoter activity maps within the DNA surrounding the hutH start codon. Pairwise comparisons of the primary structures of bacterial and mammalian histidases, together with the unique kinetic properties and gene organization, suggest that streptomycete histidase may represent a distinct family of histidases.