Post-translational modifications of protein in response to ionizing radiation

Based on central dogma of genetics, protein is the embodiment and executor of genetic function, post-translational modifications (PTMs) of protein are particularly important and involved in almost all aspects of cell biology and pathogenesis. Studies have shown that ionizing radiation (IR) alters gene expression much more profoundly and a broad variety of cell-process pathways, lots of proteins are modified and activated. Our understanding of the protein in response to ionizing radiation is steadily increasing. Among the various biological processes known to induce radioresistance, PTMs have attracted marked attention in recent years. The present review summarizes the latest knowledge about how PTMs response to ionizing radiation and pathway analysis were conducted. The data provided insights into biological effects of IR and contributing to the development of novel IR-based strategies.     

The use of a drug resistance cartridge for in vitro insertion and deletion mutagenesis of a cosmid clone

A drug-resistant cartridge was employed in the construction of families of insertion mutants of a cosmid clone. The cartridge contains a cml gene and has identical restriction enzyme sites, EcoRI, BamHI, SalI, and PstI, on both ends. The families of mutants were made by ligation of the cartridge to the cosmid, which was linearized or partially digested, followed by in vitro packaging and transduction. From these families we selected cosmid derivatives which either have a unique BamHI site at a predetermined site in the cosmid or have deletions covering different portions of the original clone. The extent of a large gene cluster cloned into the original cosmid was identified by confirming the gene function in some of the deletion mutants. The possibility for further and various uses of this cartridge is discussed.     

Role of Guanine Nucleotide Exchange Factor-H1 in Complement-mediated RhoA Activation in Glomerular Epithelial Cells

Visceral glomerular epithelial cells (GEC), also known as podocytes, are vital for the structural and functional integrity of the glomerulus. The actin cytoskeleton plays a central role in maintaining GEC morphology. In a rat model of experimental membranous nephropathy (passive Heymann nephritis (PHN)), complement C5b-9-induced proteinuria was associated with the activation of the actin regulator small GTPase, RhoA. The mechanisms of RhoA activation, however, remained unknown. In this study, we explored the role of the epithelial guanine nucleotide exchange factor, GEF-H1, in complement-induced RhoA activation. Using affinity precipitation to monitor GEF activity, we found that GEF-H1 was activated in glomeruli isolated from rats with PHN. Complement C5b-9 also induced parallel activation of GEF-H1 and RhoA in cultured GEC. In GEC in which GEF-H1 was knocked down, both basal and complement-induced RhoA activity was reduced. On the other hand, GEF-H1 knockdown augmented complement-mediated cytolysis, suggesting a role for GEF-H1 and RhoA in protecting GEC from cell death. The MEK1/2 inhibitor, U0126, and mutation of the ERK-dependent phosphorylation site (T678A) prevented complement-induced GEF-H1 activation, indicating a role for the ERK pathway. Further, complement induced GEF-H1 and microtubule accumulation in the perinuclear region. However, both the perinuclear accumulation and the activation of GEF-H1 were independent of microtubules and myosin-mediated contractility, as shown using drugs that interfere with microtubule dynamics and myosin II activity. In summary, we have identified complement-induced ERK-dependent GEF-H1 activation as the upstream mechanism of RhoA stimulation, and this pathway has a protective role against cell death.     

QUANTITATIVE VARIATION OF ECDYSTEROIDS OF IXODID TICKS

Abstract  In order to explore the role of ecdysteroids in development and reproduction of ixodid ticks, we studied the quantitative variation of ecdysteroids in the hemolymph, synganglion, ovary and whole body of the female ixodid ticks, Dermacentor niveus and Haemaphysalis longicornis , before and after engorgement and oviposition by HPLC and RIA. The ecdysteroid content in eggs of these ticks was determined by HPLC. The results indicated that before engorgement the quantitative variation of ecdysteroids in the whole female body was not significant, but their levels increased rapidly after engorgement. The ecdysteroid titer in hemolymph was peaked on the 5th day after engorgement, which was one day prior to oviposition. It may be regarded as a singnal of oviposition. In the synganglion the peak of ecdysteroid level occurred also on the 5th day after engorgement. This is coincident with the secretory activity of neurosecretory cells of synganglion. From the 3rd day after engorgement until oviposition the ecdysteroid level in the ovary increased rapidly. Ecdysteroids were detected in eggs of H. longicornis too. They stem from ovary and accumulated with the process of embryonic development.     

Influence of Phytase Transgenic Corn on the Intestinal Microflora and the Fate of Transgenic DNA and Protein in Digesta and Tissues of Broilers

An experiment was conducted to investigate the effect of phytase transgenic corn (PTC) on intestinal microflora, and the fate of transgenic DNA and protein in the digesta and tissues of broilers. A total of 160 1-day-old Arbor Acres commercial male broilers were randomly assigned to 20 cages (8 chicks per cage) with 10 cages (replicates) for each treatment. Birds were fed with a diet containing either PTC (54.0% during 1–21 days and 61.0% during 22–42 days) or non-transgenic isogenic control corn (CC) for a duration of 42 days. There were no significant differences (P>0.05) between birds fed with the PTC diets and those fed with the CC diets in the quantities of aerobic bacteria, anaerobic bacteria, colibacillus and lactobacilli, or microbial diversities in the contents of ileum and cecum. Transgenic phyA2 DNA was not detected, but phyA2 protein was detected in the digesta of duodenum and jejunum of broilers fed with the PTC diets. Both transgenic phyA2 DNA and protein fragments were not found in the digesta of the ileum and rectum, heart, liver, kidney, and breast or thigh muscles of broilers fed with the PTC diets. It was concluded that PTC had no adverse effect on the quantity and diversity of gut microorganisms; Transgenic phyA2 DNA or protein was rapidly degraded in the intestinal tract and was not transferred to the tissues of broilers.     

濒危植物永瓣藤叶片功能性状对环境因子的响应

植物叶片功能性状能够响应环境条件的变化，反应了植物对环境的适应策略。当前，针对藤本植物叶片功能性状地理格局及其环境驱动力的研究较少。以国家重点保护植物永瓣藤（Monimopetalum chinense）为研究对象，对其分布区内11个种群的15个叶片功能性状进行测量，并结合气候、土壤因子来解释叶性状变异。比较叶片性状在局域和区域尺度上的种内变异程度，利用多元逐步回归分析环境因子对叶性状的影响。结果表明，在局域尺度上，永瓣藤叶功能性状变异系数介于3.0%-22.5%，其中，叶面积变异程度最大，叶片碳含量变异最小。永瓣藤叶片形状随纬度上升而变得宽且圆。叶片磷含量相对较低，永瓣藤的生长可能受到了磷限制。土壤与气候因子是叶片性状的重要驱动因素，解释了25%-97%的叶片性状变异。在温度和水分充足的情况下，永瓣藤叶片趋向于的慢速生长的保守策略。总体来说，永瓣藤叶片功能性状通过一定的种内变异和性状组合，并与气候、土壤因子相互作用，适应当前的环境条件。     

Application of the phosphomannose-isomerase/mannose selection system in the Agrobacterium-mediated transformation of Lonicera hypoglauca Miq.

Journal of Plant Biochemistry and Biotechnology - The dried buds of Lonicera hypoglauca Miq. have antipyretic, antidotal and anti-inflammatory properties and as Flos lonicerae are widely used in...