MST1 promotes apoptosis through phosphorylation of histone H2AX

MST1 (mammalian STE20-like kinase 1) is a serine/threonine kinase that is cleaved and activated by caspases during apoptosis. Overexpression of MST1 induces apoptotic morphological changes such as chromatin condensation, but the mechanism is not clear. Here we show that MST1 induces apoptotic chromatin condensation through its phosphorylation of histone H2AX at Ser-139. During etoposide-induced apoptosis in Jurkat cells, the cleavage of MST1 directly corresponded with strong H2AX phosphorylation. In vitro kinase assay results showed that MST1 strongly phosphorylates histone H2AX. Western blot and kinase assay results with a mutant S139A H2AX confirmed that MST1 phosphorylates H2AX at Ser-139. Direct binding of MST1 and H2AX can be detected when co-expressed in HEK293 cells and was also confirmed by an endogenous immunoprecipitation study. When overexpressed in HeLa cells, both the MST1 full-length protein and the MST1 kinase domain (MST1-NT), but not the kinase-negative mutant (MST1-NT-KN), could induce obvious endogenous histone H2AX phosphorylation. The caspase-3 inhibitor benzyloxycarbonyl-DEVD-fluoromethyl ketone (Z-DEVD-fmk) attenuates phosphorylation of H2AX by MST1 but cannot inhibit MST1-NT-induced histone H2AX phosphorylation, indicating that cleaved MST1 is responsible for H2AX phosphorylation during apoptosis. Histone H2AX phosphorylation and DNA fragmentation were suppressed in MST1 knockdown Jurkat cells after etoposide treatment. Taken together, our data indicated that H2AX is a substrate of MST1, which functions to induce apoptotic chromatin condensation and DNA fragmentation.     

黏菌代谢产物及其生物活性的研究进展

黏菌代谢产物的研究显示出其较高的应用价值,并取得了较大的进展。本文综述了从黏菌中分离得到的100余个代谢产物,主要包括:脂肪酸、氨基酸、生物碱、萘醌、芳香族化合物、萜类化合物以及酯或其衍生品等。阐述了其抑菌、抗肿瘤、细胞毒及抗氧化活性,简要介绍了化合物的构效关系,同时对黏菌生物活性的研究方法及生物化学特征进行了总结,分析了不同黏菌类群代谢产物的异同。最后对黏菌代谢产物的研究提出了问题与展望。     

草莓花药培养获得无病毒植株的技术研究

以花粉发育为单核期的花药接种,在MS附加IAA 4ppm、K 2ppm、BA 2ppm的培养基上诱导愈伤组织,并可直接分化出“沙尔普斯”、“红岗利德”、“春香”3个品种的花药植株。稍加调整附加激素成分和浓度,可在“索非亚”、“宝交早生”、“戈雷拉”、“红衣”、“丽红”等品种的花药上直接经愈伤组织分化出植株。其中有“沙尔普斯”、“红岗利德”、“春香”、“宝交早生”、“索非亚”等5个品种的花药植株经过病毒检测确认不带SMoV、SCrV、SMYEV和SVBV4种病毒。对无病毒植株进行了田间对比试验,植株生长势和果实产量都明显超过对照品种。     

Theoretical Insights into Catalytic Mechanism of Protein Arginine Methyltransferase 1

Protein arginine methyltransferase 1 (PRMT1), the major arginine asymmetric dimethylation enzyme in mammals, is emerging as a potential drug target for cancer and cardiovascular disease. Understanding the catalytic mechanism of PRMT1 will facilitate inhibitor design. However, detailed mechanisms of the methyl transfer process and substrate deprotonation of PRMT1 remain unclear. In this study, we present a theoretical study on PRMT1 catalyzed arginine dimethylation by employing molecular dynamics (MD) simulation and quantum mechanics/molecular mechanics (QM/MM) calculation. Ternary complex models, composed of PRMT1, peptide substrate, and S-adenosyl-methionine (AdoMet) as cofactor, were constructed and verified by 30-ns MD simulation. The snapshots selected from the MD trajectory were applied for the QM/MM calculation. The typical S_N2-favored transition states of the first and second methyl transfers were identified from the potential energy profile. Deprotonation of substrate arginine occurs immediately after methyl transfer, and the carboxylate group of E144 acts as proton acceptor. Furthermore, natural bond orbital analysis and electrostatic potential calculation showed that E144 facilitates the charge redistribution during the reaction and reduces the energy barrier. In this study, we propose the detailed mechanism of PRMT1-catalyzed asymmetric dimethylation, which increases insight on the small-molecule effectors design, and enables further investigations into the physiological function of this family.     

Analysis of the downstream region of nodD3 P1 promoter by deletion and complementation tests in Sinorhizobium meliloti

Rhizobia specifically interacts with the host, leguminous plants, leading to the formation of nitrogen-fixing root nodules. The Rhizobium genes essential for nodule formation are called nodulation genes (nod or nol). The expression of nod genes requires the presence of host signals, generally flavonoids, and the product of regulatory nodD gene, NodD[1,2]. The expression of nod genes results in the synthesis of Nod factors, which serve as the signal molecules to elicit root cor-tical cells di…     

土壤原生动物群落及其生态功能

土壤原生动物是土壤微生物区系的重要组成部分。在土壤生态系统中 ,由于微生物与微动物的生命活动及其相互作用 ,从而形成了土壤的物质循环和能量转化。土壤原生动物既参与了微生物所介导的物质转化和能量循环 ,又参与了动物对微生物的捕食作用。由于原生动物具有丰富的种类和多样性以及巨大的生物量 ,所以土壤原生动物的群落及其生态功能 ,已引起了人们的广泛关注 ,并且研究理论与方法日益深入。但我国在这方面的研究报道较少 ,本文拟从群落与生态功能方面的进展做一概述。1　土壤原生动物的群落特征土壤与淡水原生动物最早是由Ａｎｔｏｎ…     

从野皂荚提胶后的副产物中制备分离蛋白

采用碱提酸沉法从野皂荚提胶后的副产物中制备分离蛋白,研究了制备工艺条件对分离蛋白得率的影响。结果表明:当料液比1∶10~1∶12(m/V)、提取液pH8.0~9.0、浸提温度45~50℃、浸提两次、每次浸提45 m in时,分离蛋白得率可达78.45%。研究结果可为野皂荚提胶副产物中蛋白的综合利用奠定了基础。     

中枢神经细胞的高尔基复合体ACP活性分布的电镜观察

本文采用电镜金属盐法—酸性磷酸酶(ACP)细胞化学技术,用30mmol/L pipes缓冲液配制低浓度戊二醛进行固定。对成年大鼠的大脑大锥体细胞,小脑浦肯野氏细胞,脊髓前角运动细胞的高尔基复合体的ACP活性进行了实验研究和探讨。结果发现ACP活性分布在高尔基复合体的部份转移泡、浓缩泡及GERL部位。高尔基复合体呈ACP阳性反应,并显示出多种形态。