Structure-activity relationships of Erwinia carotovora quorum sensing signaling molecules

Production of virulence factors and secondary metabolites is regulated in the phytopathogen Erwinia carotovora by quorum sensing involving N-acylated homoserine lactone (AHL) signaling molecules. Non-hydrolyzable AHL analogues were synthesized and screened in vivo. The biological activity of each compound was correlated with its ability to bind Erwinia AHL receptor proteins (LuxR homologues) in vitro. There is an excellent correlation between carbapenem production in vivo and in vitro binding to CarR. However, no such correlation could be found between exoprotease production and analogue binding to EccR. Our data are consistent with the involvement of a third, as yet uncharacterized LuxR homologue.     

Nonapoptotic functions of FADD-binding death receptors and their signaling molecules

Death receptors (DRs) are surface receptors that when triggered have the capacity to induce apoptosis in cells by forming the death-inducing signaling complex (DISC). The first protein recruited to form the DISC is the adaptor protein FADD/Mort1. Some members of the DR family, CD95 and the TRAIL receptors DR4 and DR5, directly bind FADD, whereas others, such as TNF receptor I and DR3, initially bind another adaptor protein, TRADD, which then recruits FADD. While all DRs can activate both apoptotic and non-apoptotic pathways, it has been widely assumed that the main physiological role of FADD-binding death receptors is to trigger apoptosis. However, recent work has ascribed multiple non-apoptotic activities to these receptors and/or the signaling components of the DISC.     

Marine sponges and their microbial symbionts: love and other relationships

Many marine sponges harbour dense and diverse microbial communities of considerable ecological and biotechnological importance. While the past decade has seen tremendous advances in our understanding of the phylogenetic diversity of sponge-associated microorganisms (more than 25 bacterial phyla have now been reported from sponges), it is only in the past 3-4 years that the in situ activity and function of these microbes has become a major research focus. Already the rewards of this new emphasis are evident, with genomics and experimental approaches yielding novel insights into symbiont function. Key steps in the nitrogen cycle [denitrification, anaerobic ammonium oxidation (Anammox)] have recently been demonstrated in sponges for the first time, with diverse bacteria - including the sponge-associated candidate phylum 'Poribacteria'- being implicated in these processes. In this minireview we examine recent major developments in the microbiology of sponges, and identify several research areas (e.g. biology of viruses in sponges, effects of environmental stress) that we believe are deserving of increased attention.     

Gamete membrane microdomains and their associated molecules in fertilization signaling

Fertilization is the fundamental system of biological reproduction in many organisms, including animals, plants, and algae. A growing body of knowledge has emerged to explain how fertilization and activation of development are accomplished. Studies on the molecular mechanisms of fertilization are in progress for a wide variety of multicellular organisms. In this review, we summarize recent findings and debates about the long-standing questions concerning fertilization: how egg and sperm become competent for their interaction with each other, how the binding and fusion of these gamete cells are made possible, and how the fertilized eggs initiate development to a newborn. We will focus on the structure and function of the membrane microdomains (MDs) of egg and sperm that may serve as a platform or signaling center for the aforementioned cellular functions. In particular, we provide evidence that MDs of eggs from the African clawed frog, Xenopus laevis, play a pivotal role in receiving extracellular signals from fertilizing sperm and then transmitting them to the egg cytoplasm, where the tyrosine kinase Src is present and responsible for the subsequent signaling events collectively called egg activation. The presence of a new signaling axis involving uroplakin III, an MD-associated transmembrane protein, and Src in this system will be highlighted and discussed.     

Synthesis of eperezolid-like molecules and evaluation of their antimicrobial activities

3-Fluoro-4-(4-phenylpiperazin-l-yl)aniline (II) prepared from 3,4-difluoro nitrobenzene was converted to the corresponding Schiff bases (III) and (IV) by treatment with 4-methoxybenzaldehyde and indol-3-carbaldehyde, respectively. Treatment of amine (II) with 4-fluorophenyl isothiocyanate afforded the corresponding thiourea derivative (V). Compound (V) was converted to thiazolidinone and thiazoline derivatives (VI) and (VII) by cyclocondensation with ethylbromoacetate or 4-chlorophenacylbromide, respectively. The synthesis of carbothioamide derivative (X) was performed starting from compound (II) by three steps. Treatment of compound (X) with sodium hydroxide, sulfuric acid, or chlorophenacyl bromide generated the corresponding 1,2,4-triazole (XI), 1,3,4-thiadiazole (XII), and 1,3-thiazolidinone (XIII) derivatives, respectively. The structural assignments of new compounds were based on their elemental analysis and spectral (IR, ¹H-NMR, ¹³C-NMR, and LC-MS) data. In the antimicrobial activity study all the compounds revealed high anti-Mycobacterium smegmatis activity.     

Novel cytotoxic bufadienolides derived from bufalin by microbial hydroxylation and their structure-activity relationships

Microbial transformation was used to prepare novel cytotoxic bufadienolides. Twelve products (3-14) were obtained from bufalin (1) by the fungus Mucor spinosus. Their structures were elucidated by high-resolution mass spectroscopy (HR-MS) and extensive NMR techniques, including 1H NMR, 13C NMR, DEPT, 1H-1H correlation spectroscopy (COSY), two dimensional nuclear Overhauser effect correlation spectroscopy (NOESY), heteronuclear multiple quantum coherence (HMQC), and heteronuclear multiple bond coherence (HMBC). Compounds 3, 4, 9 and 11-14 are new mono- or dihydroxylated derivatives of bufalin with novel oxyfunctionalities at C-1beta, C-7beta, C-11beta, C-12beta and C-16alpha positions. The in vitro cytotoxic activities against human cancer cell lines of 3-14, together with 16 biotransformed products derived from cinobufagin (15-30) were determined by the MTT method, and their structure-activity relationships (SAR) were discussed.     

Microbial quorum sensing signaling molecules and their roles in the biosynthesis of natural products

<正>Microorganisms act as a double-edged sword that can bring benefits and diseases to humans. Their physiological and metabolic processes are under precise control of intricate regulatory networks, in which signaling systems are critical for cells coordinating or competing with each other to overcome unfavorable environmental conditions, including the prompt and acute responses to endogenous or exogenous stimuli. In Streptomyces,     

玉米生长期间土壤微生物量与土壤酶变化及其相关性研究

研究了玉米生长期间土壤微生物量碳、氮与土壤过氧化氢酶、蔗糖酶、脲酶、蛋白酶活性变化及其相关性．结果表明，玉米生长前期和中期，土壤微生物量碳、氮及酶活性迅速上升，并逐渐达到最大值；玉米生长后期，土壤微生物量碳、氮、酶活性下降至某一值后并逐渐趋于平稳．几种处理相比较，以秸秆＋尿素处理的土壤微生物量碳、氮及酶活性为最大．除玉米生长后期，土壤微生物量碳、氮与碱解氮、活性腐殖质、土壤ｐＨ不相关外，土壤微生物量碳、氮与土壤过氧化氢酶、蔗糖酶、脲酶、蛋白酶活性及速效养分在玉米生长期间均相关或极相关     

Analysis of Ca2+ mediated signaling regulating Toxoplasma infectivity reveals complex relationships between key molecules

Host cell invasion, exit and parasite dissemination is critical to the pathogenesis of apicomplexan parasites such as Toxoplasma gondii and Plasmodium spp. These processes are regulated by intracellular Ca²⁺ signaling although the temporal dynamics of Ca²⁺ fluxes and down‐stream second messenger pathways are poorly understood. Here, we use a genetically encoded biosensor, GFP‐Calmodulin‐M13–6 (GCaMP6), to capture Ca²⁺ flux in live Toxoplasma and investigate the role of Ca²⁺ signaling in egress and motility. Our analysis determines how environmental cues and signal activation influence intracellular Ca²⁺ flux, allowing placement of effector molecules within this pathway. Importantly, we have identified key interrelationships between cGMP and Ca²⁺ signaling that are required for activation of egress and motility. Furthermore, we extend this analysis to show that the Ca²⁺ Dependent Protein Kinases–TgCDPK1 and TgCDPK3—play a role in signal quenching before egress. This work highlights the interrelationships of second messenger pathways of Toxoplasma in space and time, which is likely required for pathogenesis of all apicomplexan species.     

微生物胞外多糖的合成及其在重金属修复中的作用机制与应用

重金属的生物不可降解性使其在环境中长期存在,导致严重的环境污染,对人类健康和生态系统构成威胁。与传统的物化修复技术相比,微生物修复具有成本低廉、环境友好和高效等特点。在面对重金属胁迫或营养不均衡时,微生物会被激发以分泌合成胞外多糖(exopolysaccharides, EPS)。由此可见,EPS的产生是微生物对抗重金属胁迫的重要策略之一。EPS不仅能保护微生物在低温、高温、高盐等极端环境或受毒性化合物胁迫的条件下存活,并且在细胞内外进行信息和物质的交流与传递,既作为保护屏障限制重金属离子进入细胞,又作为介质进行交流。EPS结构中含有多个带负电荷的官能团,能够与重金属离子发生络合、离子交换、氧化还原等反应,从而降低重金属的生物有效性并减轻其毒性。微生物EPS在重金属胁迫环境中的修复具有重要意义。然而,目前缺乏关于微生物EPS合成过程、与重金属互作机制及其在重金属胁迫环境中应用现状的系统综述。本文概述了微生物EPS及其分类,详细阐述了细菌EPS胞内及胞外的生物合成机制,并探讨了微生物EPS与重金属互作机制,以及微生物EPS修复水、土环境中重金属污染方面的研究进展。最后,展望了EPS合成及其在重金属修复中的作用机制研究,可为微生物EPS进一步应用于环境重金属污染修复提供支持。     

Structure-function relationships in microbial exopolysaccharides

Sufficient well-characterized microbial exopolysaccharides are now available to permit extensive studies on the relationship between their chemical structure and their physical attributes. This is seen even in homopolysaccharides with relatively simple structures but is more marked when greater differences in structure exist, as are found in several heteropolysaccharides. The specific and sometimes unique properties have, in the case of several of these polymers, provided a range of commercial applications. The existence of "families" of structurally related polysaccharides also indicates the specific role played by certain structures and substituents; the characteristics of several of these microbial polysaccharide families will be discussed here. Thus, microbial exopolysaccharides frequently carry acyl groups which may profoundly affect their interactive properties although these groups often have relatively little effect on solution viscosity. Xanthan with or without acylation shows marked differences in synergistic gelling with plant gluco- and galacto-mannans, although the polysaccharides with different acylation patterns show similar viscosity. Similarly "gelrite" from the bacterium originally designated as Auromonas (Pseudomonas)elodea is of greater potential value after deacetylation, when it provides a valuable gelling agent, than it is as a viscosifier in the natural acylated form. The Klebsiella type 54 polysaccharide only forms gels when it, too, has been chemically deacetylated to give a structure equivalent to the Enterobacter XM6 polymer. Both these polysaccharides form gels due to the enhanced interaction with cations following deacylation and to the conformation adopted after removal of the acyl groups. Recent work in our laboratory suggests that deacetylation of certain bacterial alginates also significantly increases ion binding by these polysaccharides, making them more similar in their properties to algal alginates even although the alginates from some Pseudomonas species lack poly-L-guluronic acid sequences. The existence within families of polysaccharides of types in which monosaccharides are altered within a specific structure, or with varying side-chains, also gives an indication of the way in which such substituents affect the physical properties of the polymers in aqueous solution.     

Evolutionary relationships of microbial aromatic prenyltransferases

The linkage of isoprenoid and aromatic moieties, catalyzed by aromatic prenyltransferases (PTases), leads to an impressive diversity of primary and secondary metabolites, including important pharmaceuticals and toxins. A few years ago, a hydroxynaphthalene PTase, NphB, featuring a novel ten-stranded β-barrel fold was identified in Streptomyces sp. strain CL190. This fold, termed the PT-barrel, is formed of five tandem ααββ structural repeats and remained exclusive to the NphB family until its recent discovery in the DMATS family of indole PTases. Members of these two families exist only in fungi and bacteria, and all of them appear to catalyze the prenylation of aromatic substrates involved in secondary metabolism. Sequence comparisons using PSI-BLAST do not yield matches between these two families, suggesting that they may have converged upon the same fold independently. However, we now provide evidence for a common ancestry for the NphB and DMATS families of PTases. We also identify sequence repeats that coincide with the structural repeats in proteins belonging to these two families. Therefore we propose that the PT-barrel arose by amplification of an ancestral ααββ module. In view of their homology and their similarities in structure and function, we propose to group the NphB and DMATS families together into a single superfamily, the PT-barrel superfamily.     

Membrane lipids as signaling molecules

PURPOSE OF REVIEW: Membrane lipids play important roles in signaling reactions. They are involved in most if not all cellular signaling cascades and in a wide variety of tissue and cell types. The purpose of this review is to highlight major pathways of signaling originating in membrane lipids. Details of lipid metabolism, and its relation to protein function, will thus advance understanding of the role of lipids in health and disease. RECENT FINDINGS: Major classes of lipids including glycerophospholipids, their metabolites (eicosanoids, endocannabinoids), and sphingolipids have recently generated interest in the field of signal transduction. These lipids are tightly regulated and have an impact on various physiological functions. Importantly, aberrant lipid metabolism often leads to onset of pathology, and thus the precise balance of signaling lipids and their effectors can serve as biomarkers. SUMMARY: Membrane lipids form precursors for second messengers and functional assembly matrices on membrane domains during cellular stimulation. Many of these modifications are rapid reactions at lipid headgroups. Metabolism of the fatty acyl portion of membrane lipids leads to the generation of a bewildering complexity of lipid mediators with extended effects in space and time.     

Sugars as signaling molecules.

Recent studies indicate that, in a manner similar to classical plant hormones, sugars can act as signaling molecules that control gene expression and developmental processes in plants. Crucial evidence includes uncoupling glucose signaling from its metabolism, identification of glucose sensors, and isolation and characterization of mutants and other regulatory components in plant sugar signal transduction pathways. The emerging scenario points to the existence of a complex signaling network that interconnects transduction pathways from sugars and other hormone and nutrient signals.     

Fatty acid amide signaling molecules

Key studies leading to the discovery and definition of the role of endogenous fatty acid amide signaling molecules are summarized.     

Synthesis and structure-activity relationships of thioflavone derivatives as specific inhibitors of the ERK-MAP kinase signaling pathway

Condensation of nitrobenzaldehydes 3 and alpha-[o-(p-methoxybenzylthio)benzoyl] sulfoxide 4 gave alpha-sulfinyl enones 5. Treatment of 5 with formic acid caused cyclization followed by debenzylation to afford 3-(methylsulfinyl)thioflavanones 6. Double-bond formation with elimination of methanesulfenic acid was performed by refluxing 6 in benzene, and, finally, the nitro group of 2-phenyl-4H-1-benzothiopyran-4-one (thioflavones) 7 was reduced with tin in tetrafluoroboric acid. Various 2'-aminothioflavones 8 thus prepared were evaluated for their inhibitory effects on the ERK-MAP kinase pathway. In a cell-based assay, 2-(2'-amino-3'-methoxyphenyl)-4H-1-benzothiopyran-4-one (8b) showed a more potent inhibitory effect than the corresponding oxygen compound (PD98059, 1) on the Raf-induced activation of the ERK-MAP kinase pathway as well as cell proliferation. Furthermore, compound 8b selectively and potently inhibited the proliferation of tumor cells in which the ERK-MAP kinase pathway is constitutively activated.     

杉木人工林土壤微生物生物量碳氮特征及其与土壤养分的关系

研究了湖南会同红黄壤区杉木人工林和常绿阔叶林土壤微生物量和养分状况.结果表明,该区杉木人工林取代地带性常绿阔叶林和杉木连栽后,土壤微生物碳、氮和土壤养分含量下降,土壤严重退化.在0～10 cm土层内,常绿阔叶林土壤微生物碳和氮含量为800.5和84.5 mg·kg^-1,分别是第1代杉木林的1.90和1.03倍、第2代杉木林的2.16和1.27倍;在10～20 cm土层内,常绿阔叶林土壤微生物碳和氮含量为475.4和63.3 mg·kg^-1,分别是第1代杉木纯林的1.86、1.60倍和第2代杉木林的2.11和1.76倍.在0～10 cm 和10～20cm土层内,杉木人工林取代常绿阔叶林和杉木栽植代数增加后,土壤全氮、全钾、铵态氮和速效钾含量均明显降低,但差异并不显著.人工杉木林林分组成单一,其凋落物分解慢、归还养分数量少;炼山等造成的表土流失是杉木人工林土壤微生物量和养分库退化的重要原因.土壤微生物碳与土壤全氮、铵态氮、全钾和速效钾含量呈极显著的正相关,土壤微生物氮与土壤养分含量也达到极显著水平.     

Synthesis of antisense oligonucleotides carrying modified 2-5A molecules at their 5'-termini and their properties

The synthesis of 8-methyladenosine-substituted 2-5A tetramers with hydroxyalkyl groups at the 5'-phosphates and the corresponding 2-5A-antisense chimeras is described. These oligonucleotides were synthesized by the phosphoramidite method with a DNA/RNA synthesizer. These 2-5A tetramers with hydroxyethyl and hydroxybutyl groups at their 5'-phosphates were more resistant to hydrolysis by alkaline phosphatase than those without the hydroxyalkyl groups. Incorporation of the hydroxyethyl group into the 2-5A tetramer and 2-5A-antisense chimera slightly reduced the abilities of their analogues to activate recombinant human RNase L, but the abilities of the 2-5A tetramer and the 2-5A-antisense chimera both with the hydroxyethyl group and 8-methyladenosine returned to 80 and 50% relative to those of the oligonucleotides without the hydroxyethyl group and 8-methyladenosine, respectively. Furthermore, the enzyme activated by 8-methyladenosine-substituted 2-5A-antisense chimera with the hydroxyethyl group cleaved the complementary RNA as efficiently as that activated by 2-5A-antisense chimera without the hydroxyethyl group and 8-methyladenosine. Thus, the 2-5A-antisense chimera carrying the hydroxyethyl group and 8-methyladenosine will be a candidate for a novel antisense molecule.