Movement-associated proteins of Potato virus A: attachment to virus particles and phosphorylation

The particles of Potato virus A (PVA; genus Potyvirus) are helically constructed filaments that contain multiple copies of a single type of coat-protein (CP) subunit and a single copy of genome-linked protein (VPg), attached to one end of the virion. Examination of negatively-stained virions by electron microscopy revealed flexuous, rod-shaped particles with no obvious terminal structures. It is known that particles of several filamentous plant viruses incorporate additional minor protein components, forming stable complexes that mediate particle disassembly, movement or transmission by insect vectors. The first objective of this work was to study the interaction of PVA movement-associated proteins with virus particles and how these interactions contribute to the morphology and function of the virus particles. Purified particles of PVA were examined by atomic force microscopy (AFM) and immuno-gold electron microscopy. A protrusion was found at one end of some of the potyvirus particles, associated with the 5' end of the viral RNA. The tip contained two virus-encoded proteins, the genome-linked protein (VPg) and the helper-component proteinase (HC-Pro). Both are required for cell-to-cell movement of the virus. Biochemical and electron microscopy studies of purified PVA samples also revealed the presence of another protein required for cell-to-cell movement the cylindrical inclusion protein (CI), which is also an RNA helicase/ATPase. Centrifugation through a 5-40% sucrose gradient separated virus particles with no detectable CI to a fraction that remained in the gradient, from the CI-associated particles that went to the pellet. Both types of particles were infectious. AFM and translation experiments demonstrated that when the viral CI was not present in the sample, PVA virions had a beads-on-a-string phenotype, and RNA within the virus particles was more accessible to translation. The second objective of this work was to study phosphorylation of PVA movement-associated and structural proteins (CP and VPg) in vitro and, if possible, in vivo. PVA virion structural protein CP is necessary for virus cell-to-cell movement. The tobacco protein kinase CK2 was identified as a kinase phosphorylating PVA CP. A major site of CK2 phosphorylation in PVA CP was identified as a single threonine within a CK2 consensus sequence. Amino acid substitutions affecting the CK2 consensus sequence in CP resulted in viruses that were defective in cell-to-cell and long-distance movement. The CK2 regulation of virion assembly and cell-to-cell movement by phosphorylation of CP was possibly due to the inhibition of CP binding to viral RNA. Four putative phosphorylation sites were identified from an in vitro phosphorylated recombinant VPg. All four were mutated and the spread of mutant viruses in two different host plants was studied. Two putative phosphorylation site mutants (Thr45 and Thr49) had phenotypes identical to that of a wild type (WT) virus infection in both Nicotiana benthamiana and N. tabacum plants. The other two mutant viruses (Thr132/Ser133 and Thr168) showed different phenotypes with increased or decreased accumulation rates, respectively, in inoculated and the first two systemically infected leaves of N. benthamiana. The same mutants were occasionally restricted to single cells in N. tabacum plants, suggesting the importance of these amino acids in the PVA infection cycle in N. tabacum.

Unpacking the impact of attachment to project teams on boundary-spanning behaviors

As business environments become even more competitive, project teams are required to make an effort to operate external linkages from within an organization or across organizational boundaries. Nevertheless, some members boundary-span less extensively, isolating themselves and their project teams from external environments. Our study examines why some members boundary-span more or less through the framework of group attachment theory. Data from 521 project-team members in construction and engineering industries revealed that the more individuals worry about their project team’s acceptance (group attachment anxiety), the more likely they are to perceive intergroup competition, and thus put more efforts into operating external linkages and resources to help their own teams outperform competitors. In contrast, a tendency to distrust their project teams (group attachment avoidance) generates members’ negative construal of their team’s external image, and thus fewer efforts are made at operating external linkages. Thus, project leaders and members with high group-attachment-anxiety may be best qualified for external tasks.

Role of cell attachment in leaching of chalcopyrite mineral by Thiobacillus ferrooxidans

Experiments on the leaching of copper from chalcopyrite mineral by the bacterium Thiobacillus ferrooxidans show that, in the presence of adequate amounts of sulphide, iron-grown bacteria preferentially oxidise sulphur in the ore (through direct attachment) rather than ferrous sulphate in solution. At 20% pulp density, the leaching initially takes place by a predominantly direct mechanism. The cell density in the liquid phase increases, but the Fe2+ is not oxidised. However, in the later stages when less solid substrate is available and the cell density becomes very high, the bacteria start oxidising Fe2+ in the liquid phase, thus contributing to the indirect mechanism of leaching. Contrary to expectations, the rate of leaching increased with increasing particle size in spite of the decreasing specific surface area. This has been found to be due to increasing attachment efficiency with increase in particle size.

The effects of community attachment and information seeking on displaced disaster victims’ decision making

This paper uses original survey data of the Great East Japan earthquake disaster victims to examine their decision to apply for the temporary housing as well as the timing of application. We assess the effects of victims’ attachment to their locality as well as variation in victims’ information seeking behavior. We additionally consider various factors such as income, age, employment and family structure that are generally considered to affect the decision to choose temporary housing as victims’ solution for their displacement. Empirical results indicate that, ceteris paribus, as the degree of attachment increases, victims are more likely to apply for the temporary housing but attachment does not affect the timing of application. On the other hand, the victims who actively seek information and are able to collect higher quality information are less likely to apply for the temporary housing and if they do apply then they apply relatively later.

A split cassette attachment for recording all the reflections on the upper level Weissenberg photographs either as elongated or as contracted spots

A split-cassette arrangement has been incorporated in the Weissenberg camera for recording all reflections on the upper level photographs either as elongated or as contracted spots. This arrangement employs two semicylindrical cassettes which are separated by a horizontal plane. These half-cassettes are translated in opposite directions. A suitable split-cassette attachment has been constructed for the Unicam Weissenberg goniometer S-35 The subject of 'displaced-film' Weissenberg photograph is also discussed.

Hydrogel-assisted synthesis of nanotubes and nanorods of US, ZnS and CuS, showing some evidence for oriented attachment

By carrying out the reaction of appropriate metal compounds with Na2S in the presence of a tripodal cholamide-based hydrogel, nanotubes and nanorods of CdS, ZnS and CuS have been obtained. The nanostructures have been characterized by transmission electron microscopy and spectroscopic techniques. Evidence is presented for the assembly of short nanorods to form one-dimensional chains.

Surface Ligand Population Controlled Oriented Attachment: A Case of CdS Nanowires

using two types of organic ligands having similar chemical structure but different physical properties and varying their dynamic population at the surface of zinc blende seed nanocrystals, self-assembled zinc blende semicircular-shaped bent nanowires of CdS are synthesized via a colloidal synthetic approach. It is found that the hydrophobic tail interaction of long-chain ligands puts strain on these thin nanowires (< 2 nm diameter) and bend them to some extent, forming strained nanowires.

Covalent attachment of two important dicopper(II) systems in a one-dimensional polymeric chain: An x-ray structure of [Cu2(en)2(OH)2·Cu2(O2CMe)4](PF6)2·0.5EtOH]α showing the shortest Cucdots, three dots, centeredCu Distance in the tetraacetato core

Reaction of Cu2(O2CMe)4(H2O)2 with 1,2-diaminoethane(en) in ethanol, followed by the addition of NH4PF6, led to the formation of a covalently linked 1D polymeric copper(II) title complex showing alternating [Cu2(en)2(OH)22+] and [Cu2(O2CMe)4] units in the chain and the shortest Cucdots, three dots, centeredCu separation of 2.558(2) Å in the tetraacetato core.

Exploiting oriented attachment in stabilizing La3+-doped gallium oxide nano-spindles

Retaining the morphology of gallium oxide nanostructures during structural transformations or after doping with lanthanide ions is not facile. Here we report on the sonochemical synthesis of nearly monodisperse similar to 550 nm long nano-spindles of undoped and La-doped alpha-GaOOH. The transformation of as-prepared undoped and La-doped alpha-GaOOH powders into the corresponding undoped and La-doped Ga2O3 phases (alpha and beta) was achieved by carrying out controlled annealing at elevated temperatures under optimized conditions. The formation of gallium oxide nano-spindles is explained by invoking the phenomenon of oriented attachment, as amply supported by electron microscopy. Interestingly, the morphology of the gallium oxide nano-spindles remained conserved even after doping them with more than 1.4 at% of La3+ ions. Such robust structural stability could be attributed to the oriented attachment-type growth observed in the nano-spindles. The as-prepared samples and the corresponding annealed ones were thoroughly characterized by powder X-ray diffraction (PXRD), electron microscopy (SEM, TEM, and STEM-EDS) and X-ray photoelectron spectroscopy (XPS). Finally, photoluminescence from the single-crystalline undoped and La-doped beta-Ga2O3 was explored.

Arc Root Attachment on the Anode Surface of Arc Plasma Torch Observed with a Novel Method

The arc-root attachment on the anode surface of a dc non-transferred arc plasma torch has been successfully observed using a novel approach. A specially designed copper mirror with a boron nitride film coated on its surface central-region is employed to avoid the effect of intensive light emitted from the arc column upon the observation of weakly luminous arc root. It is found that the arc-root attachment is diffusive on the anode surface of the argon plasma torch, while constricted arc roots often occur when hydrogen or nitrogen is added into argon as the plasma-forming gas.

Surface attachment of protein fibrils via covalent modification strategies.

Chemical control of surface functionality and topography is an essential requirement for many technological purposes. In particular, the covalent attachment of monomeric proteins to surfaces has been the object of intense studies in recent years, for applications as varied as electrochemistry, immuno-sensing, and the production of biocompatible coatings. Little is known, however, about the characteristics and requirements underlying surface attachment of supramolecular protein nanostructures. Amyloid fibrils formed by the self-assembly of peptide and protein molecules represent one important class of such structures. These highly organized beta-sheet-rich assemblies are a hallmark of a range of neurodegenerative disorders, including Alzheimer's disease and type II diabetes, but recent findings suggest that they have much broader significance, potentially representing the global free energy minima of the energy landscapes of proteins and having potential applications in material science. In this paper, we describe strategies for attaching amyloid fibrils formed from different proteins to gold surfaces under different solution conditions. Our methods involve the reaction of sulfur containing small molecules (cystamine and 2-iminothiolane) with the amyloid fibrils, enabling their covalent linkage to gold surfaces. We demonstrate that irreversible attachment using these approaches makes possible quantitative analysis of experiments using biosensor techniques, such as quartz crystal microbalance (QCM) assays that are revolutionizing our understanding of the mechanisms of amyloid growth and the factors that determine its kinetic behavior. Moreover, our results shed light on the nature and relative importance of covalent versus noncovalent forces acting on protein superstructures at metal surfaces.