Activities of the peptidyl transferase center of ribosomes lacking protein L27

The ribosome is the molecular machine responsible for protein synthesis in all living organisms. Its catalytic core, the peptidyl transferase center (PTC), is built of rRNA, although several proteins reach close to the inner rRNA shell. In the Escherichia coli ribosome, the flexible N-terminal tail of the ribosomal protein L27 contacts the A- and P-site tRNA. Based on computer simulations of the PTC and on previous biochemical evidence, the N-terminal α-amino group of L27 was suggested to take part in the peptidyl-transfer reaction. However, the contribution of this group to catalysis has not been tested experimentally. Here we investigate the role of L27 in peptide-bond formation using fast kinetics approaches. We show that the rate of peptide-bond formation at physiological pH, both with aminoacyl-tRNA or with the substrate analog puromycin, is independent of the presence of L27; furthermore, translation of natural mRNAs is only marginally affected in the absence of L27. The pH dependence of the puromycin reaction is unaltered in the absence of L27, indicating that the N-terminal α-amine is not the ionizing group taking part in catalysis. Likewise, L27 is not required for the peptidyl-tRNA hydrolysis during termination. Thus, apart from the known effect on subunit association, which most likely explains the phenotype of the deletion strains, L27 does not appear to be a key player in the core mechanism of peptide-bond formation on the ribosome.     

Growth regulation, plastid differentiation and the development of a photosynthetic system in cultured carrot root explants as influenced by exogenous sucrose and various phytohormones

Chromoplasts, which exist in the cells of freshly isolated carrot root explants, seemed to be transformed in thylakoid containing plastids, and chlorophyll formation was initiated if the explants were cultured in a liquid medium containing inositol and IAA as a hormonal supplement. This process was intensified when kinetin was also added, but no dependence on a sucrose supply could be found.A sucrose supply of 2% in conjunction with the combination of all three hormones, however, was needed to achieve maximal thylakoid formation including stacking in individual chloroplasts and for the very extensive chloroplast multiplication in explants growing with high cell division activity. It should be noted that the number of plastids per cell is strongly increased by the sucrose supplement which leads also to starch accumulation. However, no transformation into chloroplasts occurred without the hormonal stimulus.     

Dictyostelium Sun-1 connects the centrosome to chromatin and ensures genome stability

The centrosome-nucleus attachment is a prerequisite for faithful chromosome segregation during mitosis. We addressed the function of the nuclear envelope (NE) protein Sun-1 in centrosome-nucleus connection and the maintenance of genome stability in Dictyostelium discoideum . We provide evidence that Sun-1 requires direct chromatin binding for its inner nuclear membrane targeting. Truncation of the cryptic N-terminal chromatin-binding domain of Sun-1 induces dramatic separation of the inner from the outer nuclear membrane and deformations in nuclear morphology, which are also observed using a Sun-1 RNAi construct. Thus, chromatin binding of Sun-1 defines the integrity of the nuclear architecture. In addition to its role as a NE scaffold, we find that abrogation of the chromatin binding of Sun-1 dissociates the centrosome-nucleus connection, demonstrating that Sun-1 provides an essential link between the chromatin and the centrosome. Moreover, loss of the centrosome-nucleus connection causes severe centrosome hyperamplification and defective spindle formation, which enhances aneuploidy and cell death significantly. We highlight an important new aspect for Sun-1 in coupling the centrosome and nuclear division during mitosis to ensure faithful chromosome segregation.     

Characterization of Receptors for Substance P in Human Astrocytoma Cells: Radioligand Binding and Inositol Phosphate Formation

UC11 cells, derived from a human astrocytoma, have a high density of functional substance P receptors. Radioligand binding studies were conducted with the highly selective neurokinin-1 receptor ligand [3H][Sar9,Met(O2)11]-substance P. Kinetic binding experiments conducted at 4 degrees C yielded an association rate constant k1 of 1.86 x 10(7) M-1 min-1, a dissociation rate constant k-1 of 0.00478 min-1, and a calculated kinetic KD of 257 pM. Saturation binding experiments yielded average values of KD = 447 +/- 103 pM, Bmax = 862 +/- 93 fmol/mg of protein. This Bmax corresponds to more than 150,000 binding sites/cell. Competition binding experiments with unlabeled [Sar9,Met(O2)11]-substance P yielded average values of KD = 491 +/- 48 pM and Bmax = 912 +/- 67 fmol/mg of protein. In [3H]inositol-labeled cells, substance P induced a robust inositol phosphate formation. Inositol trisphosphate levels increased as much as 20-fold within approximately 15 s of addition of substance P. This inositol trisphosphate formation was transient and had returned to baseline within the first 60-120 s. Inositol monophosphate formation, however, was linear for at least 2 h. Structure activity data on binding and inositol monophosphate formation confirmed the presence of a neurokinin-1 receptor subtype in these cells. Thus, the UC11 cell should be a useful model cell for delineating the physiological role of substance P receptors in astrocytes.     

开花结实期小麦氮素损失的初步研究

在气候箱试验条件下,用密闭箱法研究了开花结实期小麦植株NH3、N2O、NO和NO2挥发特征及规律.结果表明:(1)只保留小麦根系处理有明显挥发NH3的作用,但在气候箱控制、白昼有相对良好的光照条件下,小麦地上部没有显著净挥发NH3作用效果;(2)开花结实期小麦植株有明显从空气吸收NO和NO2的效应,NO净吸收速率在0.5～2.7 ug·pot-1·h-1之间,NO2净吸收速率约0.4～1.6 ug·pot-1·h-1;(3)小麦根系是产生N2O的重要部位,只保留小麦根系和保留完整小麦植株处理均有显著净挥发N2O的作用效果.研究发现,开花结实期小麦植株氮素挥发损失是以N2O为主而不足NH3;这一时期小麦植株有净吸收NO、NO2明显效应,但其吸收速率远低于小麦植株N2O排放速率;开花结实期小麦N2O排放速率基本可以表征其氮素挥发损失数量.     

Post-Infection Development and Histopathology of Meloidogyne incognita in Resistant Cotton

The numbers of Meloidogyne incognita larvae which migrated from cotton roots declined over a 16-day period, but the difference in numbers migrating from resistant and susceptible cultivars was not significant. Larvae penetrated susceptible roots, matured, and reproduced within 14 days following inoculation, whereas nematode development in the resistant roots was greatly retarded. Three types of histological responses were observed in infected, resistant roots, and these correlated with the degree of nematode development. Some galls were examined which contained only fragments of nematodes; others contained no detectable traces of developing larvae. Formation of druses in galls, but not in healthy tissue, was noted in both cultivars 20 days after inoculation. Massive invasion of roots resulted in deep longitudinal fissures of root cortex.     

基于植物功能群的生态系统服务形成与维持机制研究

生态系统可以从“结构-功能-服务”3个层次来理解,其中服务是人类的主观感受或效用。维持高质量的生态系统服务还需从生态系统的自然属性入手。结构和功能是生态系统服务形成和维持的内在机制,而植物是生态系统结构和功能的核心驱动力。植物功能群具有特定功能的植物组合,其中优势植物功能群控制着生态系统的结构和功能。生态系统服务的概念、分类与植物功能群密切相关,植物功能群是生态系统与生态系统服务间更直接的桥梁。建立生态系统服务与植物功能群间的联系,使相关研究有更明确的指向,在深化理论研究的同时使研究成果更容易落地。生态系统服务的形成、维持机制与植物功能群的内在联系主要体现在4个主要方面：(1)植物功能群的内在适应性特征和外在效应是生态系统服务形成的基础;(2)植物功能群的多功能特性为生态系统服务协同提供了可能;(3)植物功能群间替代和互补效应可以提升生态系统服务质量和稳定性;(4)植物功能群能够提供基于自然的生态系统服务修复问题解决方案。     

Interaction of nematodes with nematophagus fungi: induction of trap formation, attraction and detection of attractants

Induction of trap formation in Arthrobotrys oligospora, A. conoides and Monacrosporium cystosporum was studied during a 24 h period in the presence of the free-living nematodes Panagrellus redivivus on various nutrient media. A definite pattern of trap formation was observed in these fungi. Low nutrient mineral salt medium had the most pronounced effect on trap formation. Attraction and repulsion of P. redivivus towards these three nematophagous and three non-nematophagous fungi was studied. The nematodes were attracted towards three nematophagous and one non-nematophagous fungi, the other two repelled them. Attractants of the nematophagous fungi were determined by thin-layer chromatography. In the case of A. oligospora and M. cystosporum, four spots were traced, whereas in A. conoides, five spots were detected.     

Intergeneric complementation by Agrobacterium tumefaciens chromosomal genes and its potential use for linkage mapping

The IncP1 type plasmids pULB113 (RP4:: mini-Mu) and pJB3JI were used for chromosome mobilization and R prime formation in Agrobacterium tumefaciens. With pULB113, R primes were selected for complementation of an auxotrophic marker in an Alcaligenes eutrophus recipient strain. With pJB3JI, R primes containing the Agrobacterium chromosomal virulence region chv, inactivated by a Tn5 insertion, were constructed. Selection was for kanamycin resistance in E. coli strain GV1000. Complementation tests were performed in auxotrophic A. tumefaciens, A. eutrophus and Pseudomonas fluorescens strains. This allowed us to map 9 loci, including the chv loci, in a region accounting for about 10% of the Agrobacterium chromosome.