Regulation of Arabidopsis cryptochrome 2 by blue-light-dependent phosphorylation

Cryptochromes are blue/ultraviolet-A light receptors that mediate various light responses in plants and animals. But the initial photochemical reaction of cryptochrome is still unclear. For example, although most photoreceptors are known to undergo light-dependent protein modification such as phosphorylation, no blue-light dependent phosphorylation has been reported for a cryptochrome. Arabidopsis cryptochrome 2 (cry2) mediates light regulation of seedling development and photoperiodic flowering. The physiological activity and cellular level of cry2 protein are light-dependent, and protein protein interactions are important for cry2 function. Here we report that cry2 undergoes a blue-light-dependent phosphorylation, and that cry2 phosphorylation is associated with its function and regulation. Our results suggest that, in the absence of light, cry2 remains unphosphorylated, inactive and stable; absorption of blue light induces the phosphorylation of cry2, triggering photomorphogenic responses and eventually degradation of the photoreceptor.     

Regulation of pro-opiomelanocortin biosynthesis and processing by transplantation immunity

It is more than thirty years since Billingham and Medawar showed that adrenocorticotrophic hormone (ACTH) and cortisol can prolong the survival of skin allografts. It has since become clear that glucocorticoid hormones are critically involved in the regulation of immunity. The level of glucocorticoids secreted in response to antigenic challenge corresponds to the magnitude of the immune response and in general reaches immunosuppressive levels. Interestingly, not all immune responses enhance ACTH and glucocorticoid hormone production. In transplantation immunity, the reverse seems to be true: circulating glucocorticoid levels at the time of skin graft rejection are lower than control levels. Because beta-endorphin and ACTH originate from the same prohormone, pro-opiomelanocortin (POMC), and are closely related in their tissue-specific processing and coordinate release, we have investigated the role of pituitary beta-endorphin in transplantation immunity. We report here that POMC biosynthesis and processing in the pars intermedia, but not in the anterior pituitary, can be regulated by T cell-specific factors secreted in animals undergoing transplantation immunity.     

拟南芥VHA-c基因的矿质营养调节

对Ca2+和MS营养处理转VHA-c-GUS烟草叶片的GUS表达活性进行分析。研究结果表明:VHA-c2和VHA-c3可被Ca2+促进表达,而VHA-c5的表达则被Ca2+抑制。同时,MS营养成分可显著促进VHA-c2、VHA-c3和VHA-c5基因的表达。这说明VHA-c基因可被Ca2+和MS等营养成分调控表达。     

Regulation of glutamate receptor binding by the cytoskeletal protein fodrin

The erythrocyte cytoskeleton, which consists primarily of a meshwork of spectrin and actin, controls cell shape and the disposition of proteins within the membrane. Proteins similar to spectrin have recently been found in diverse cells and tissues, and it is possible that they mediate the capping of cell-surface receptors, although this has not been demonstrated directly. In neurones, the spectrin-like protein fodrin lines the cortical cytoplasm and may link actin filaments to the membrane. Fodrin has been hypothesized to regulate the number of receptor binding sites on neuronal membranes for the putative neurotransmitter L-glutamate. Micromolar calcium concentrations activate the thiol protease calpain I, induce fodrin degradation and more than double the density of glutamate binding sites; these effects are all blocked by thiol protease inhibitors. We have now used specific antibodies to examine further the role of fodrin proteolysis in regulating glutamate receptors. We report that fodrin antibodies block the fodrin degradation and increase in glutamate binding normally induced by calcium, and so provide direct evidence for control of membrane receptors by a non-erythroid spectrin.     

Deg1 is involved in the degradation of the PsbO oxygen-evolving protein of photosystem II in Arabidopsis

Deg1, a thylakoid lumen-localized protease, retains both chaperone and protease activities. The in vivo function of Deg1 has been shown to be involved not only in PSII assembly but also in the degradation of PSII reaction center protein D1. Here we used the transgenic plants with reduced Deg1 to examine whether the lumen-localized proteins are also the substrates of Deg1 in vivo. Our results showed that the transgenic plants accumulated degradation products of the PsbO protein while the levels of full-length PsbO were not affected. The PsbO degradation products could be efficiently degraded by the recombinant Deg1. These results suggest that Deg1 is involved in the degradation of the PsbO degradation fragments, but not in the initial cleavage event itself.     

Vernalization in Arabidopsis thaliana is mediated by the PHD finger protein VIN3

In biennials and winter annuals, flowering is typically blocked in the first growing season. Exposure to the prolonged cold of winter, through a process called vernalization, is required to alleviate this block and permit flowering in the second growing season. In winter-annual types of Arabidopsis thaliana, a flowering repressor, FLOWERING LOCUS C (FLC), is expressed at levels that inhibit flowering in the first growing season. Vernalization promotes flowering by causing a repression of FLC that is mitotically stable after return to warm growing conditions. Here we identify a gene with a function in the measurement of the duration of cold exposure and in the establishment of the vernalized state. We show that this silencing involves changes in the modification of histones in FLC chromatin.     

Regulation of endothelium-derived nitric oxide production by the protein kinase Akt.

Endothelial nitric oxide synthase (eNOS) is the nitric oxide synthase isoform responsible for maintaining systemic blood pressure, vascular remodelling and angiogenesis. eNOS is phosphorylated in response to various forms of cellular stimulation, but the role of phosphorylation in the regulation of nitric oxide (NO) production and the kinase(s) responsible are not known. Here we show that the serine/threonine protein kinase Akt (protein kinase B) can directly phosphorylate eNOS on serine 1179 and activate the enzyme, leading to NO production, whereas mutant eNOS (S1179A) is resistant to phosphorylation and activation by Akt. Moreover, using adenovirus-mediated gene transfer, activated Akt increases basal NO release from endothelial cells, and activation-deficient Akt attenuates NO production stimulated by vascular endothelial growth factor. Thus, eNOS is a newly described Akt substrate linking signal transduction by Akt to the release of the gaseous second messenger NO.     

PREPARATIOPREPARATION OF 1-OCTENE BY ETHYLENE TETRAMERIZATION WITH HIGH SELECTIVITY

1-OCTENE IS WIDELY USED IN THE CHEMICAL INDUSTRY AS A COMONOMER FOR LINEAR LOW-DENSITY POLYETHYLENE (LLDPE), PLASTICIZERS, DETERGENT ALCOHOLS AND SYNTHETIC LUBRICANTS[1]. THE ETHYLENE COPOLYMER WITH 1-OCTENE AS COMONOMER HAS BETTER MECHANICAL PROPERTIES, …     

Preparation of 1-octene by ethylene tetramerization with high selectivity

Two novel PNP ligands have been synthesized and characterized by ^1H-NMR, elemental analysis, and mass spectra. In combination with Cr（Ⅲ） and cocatalyst MAO, they generate active catalytic systems that tetramerize ethylene with both high catalytic activity and high selectivity to produce 1-octene. The results show that these catalyst systems are able to catalyze ethylene tetramerization, with high catalytic activity up to 0.89×10^6 g/mol Cr.h, and the selectivity of C8 in products is 72.52%, and the percentage of 1-olefins in the C8 cut is 97.87%.     

拟南芥AtERF1蛋白的原核可溶性表达及多克隆抗体的制备

转录因子AtERF1属于ERF/AP2家族的B3亚家族,参与植物的防卫反应.根据密码子简并原理,用基因全合成的方法合成了满足大肠杆菌密码子嗜好的编码拟南芥AtERF1蛋白的碱基序列,并将其克隆到原核表达载体pET-29b.将表达载体AtERF1-pET-29b转入大肠杆菌BL21(DE3),用IPTG成功诱导表达了分子量约30kD的AtERF1蛋白,该蛋白主要以可溶性蛋白的形式存在.以该蛋白为抗原免疫家兔,制备出的多克隆抗体经免疫双扩散测定效价达1∶16,Western blot检测表明该抗血清与AtERF1蛋白识别良好.AtERF1抗体的制备为进一步从生化水平上研究AtERF1的功能奠定了良好基础.     

HIF-1调控小鼠bace1基因的转录表达

阿尔茨海默症的一个关键致病原因是大脑中淀粉样蛋白（A13）的过度产生或堆积．Aβ是由其前体蛋白APP经β-和γ-分泌酶依次水解而生成的，但对于这些分泌酶基因表达的转录调控的了解还很少．由于大脑发生缺氧／缺血会造成Aβ的产量增加，而缺氧时所激活的转录因子HIF-1（Hypoxia-Inducible Factor 1）会调控下游多种基因的表达，我们对HIF-1是否参与调控β-分泌酶的表达进行了研究．对小鼠β-分泌酶基因bace1的调控序列进行分析，发现其中含有一个缺氧应答元件（Hypoxia-Responsive Element，HRE）．我们的数据显示，HRE突变片段启动报告基因荧光素酶表达的活性比正常序列片段的启动活性明显降低．电泳迁移率的变动分析（EMSA）实验进一步证实，HIF-1可以与小鼠bace1中HRE元件相互作用．当在哺乳动物细胞中过度表达HIF-1时，BACE1的mRNA水平和蛋白质水平均有明显的增高．这些结果表明小鼠bace1基因的表达受转录因子HIF-1的调控．鉴于目前阿尔茨海默症研究领域都把抑制BACE1作为首选治疗靶位，HIF-1因而有可能成为治疗AD的一种药物靶点．     

气相色谱法测定卷烟主流烟气气相物中的甲烷、乙烯和乙炔

 建立了测定卷烟主流烟气气相物中的甲烷、乙烯和乙炔气体的气相色谱法,用外标法定量,同时考查了捕集条件和仪器的稳定性.所建立的方法简便、快速、准确,并具有良好的线性关系(r_甲烷²=0.9993,r_乙烯²=0.9998,r_乙炔²=0.9994)及较好的准确度与精密度,回收率均>85%,相对标准偏差<3%.将该方法应用于实际样品检测,结果令人满意.