The Core Histone-binding Region of the Murine Cytomegalovirus 89K Immediate Early Protein

The gene regulatory immediate early protein, pp89, of murine cytomegalovirus interacts with both DNA-associated and isolated histones in vitro. We characterized the histone-binding region of pp89 and its cellular localization during cell division to examine the possible interaction between pp89 and chromatin. pp89 expressed constitutively in cell line BALB/c 3T3 IE1 does not interact with condensed chromatin. As observed in infected cells, pp89 is localized within the nucleus of cells during interphase but spreads throughout the cell plasma following degradation of the nuclear membrane during early mitosis. In late telophase, pp89 is reorganized within the nucleus. Analysis of pp89 deletion mutants and of fragments generated by cleavage at pH 2·5 revealed that the regions responsible for association with histone are located between amino acids 71 and 415, and are not identical with the domain that shows homology to histone H2B or the highly acidic carboxy-terminal region. A potential gene-activating role of the high affinity of pp89 for isolated histones and the low affinity for DNA-associated histones is discussed

An acidic region of the 89K murine cytomegalovirus immediate early protein interacts with DNA

The product of the ie 1 gene, the regulatory immediate early protein pp89 of murine cytomegalovirus (MCMV), interacts with core histones, which can mediate the association of pp89 with DNA. We report the capacity of pp89 to interact directly with DNA in the absence of cellular proteins. After separation of proteins by SDS–PAGe, pp89 bound ds- and ssDNA, with a preference for ssDNA. Binding to specific DNA sequences in the MCMV genome was not detected. The DNA-binding region of pp89 was located to amino acids 438 to 534 by analysis of deletion mutants expressed as -galactosidase or TrpE fusion proteins. This region is identical to the highly acidic C-terminal region spanning amino acids 424 to 532. The human cytomegalovirus IE1 protein, which contains a similar extended C-terminal acidic region, does not react with DNA under the same experimental conditions

Impact of inundation regime on wild bee assemblages and associated bee–flower networks

International audienceAbstractWild bee assemblages on flood-prone meadows were compared with those on rarely inundated sites along the river Danube in easternmost Lower Austria. We sampled flower-visiting bees on 32 meadows from April to August 2016. Although we recorded more bee individuals on rarely inundated meadows, total bee species richness was higher on regularly flooded meadows and we observed a stronger differentiation diversity of bees among annually flooded meadows. Three network metrics derived from a bipartite plant–bee interaction matrix were unaffected by flooding regime. We conclude that extreme floods, which sporadically affect the investigated habitats, may have a devastating effect on wild bee populations, but communities quickly recover. This resilience surely depends on recolonization from the surrounding landscape, which emphasizes the need to consider community dynamics in highly variable floodplain areas not only locally, but on a landscape scale

The 89,000-Mr murine cytomegalovirus immediate-early protein activates gene transcription

To study trans-activation of gene expression by murine cytomegalovirus (MCMV) immediate-early (IE) proteins, the IE coding region 1 (ie1), which encodes the 89,000-Mr IE phosphoprotein (pp89), was stably introduced into L cells. A cell line was selected and characterized that efficiently expressed the authentic viral protein. The pp89 that was constitutively expressed in L cells stimulated the expression of transfected recombinant constructs containing the bacterial chloramphenicol acetyltransferase (CAT) gene under the control of viral promoters. The regulatory function of the ie1 product was confirmed by transient expression assays in which MCMV IE genes were cotransfected into L cells together with recombinant constructs of the CAT gene. For CAT activation by the ie1 product, a promoter region was required, but there was no preferential activation of a herpes simplex virus type 1 delayed-early promoter. All plasmid constructs that contained the intact coding sequences for pp89 induced gene expression in trans. The MCMV enhancer region was not essential for the expression of a functional IE gene product, and testing of the cis-regulatory activity of the MCMV enhancer revealed a low activity in L cells. Another region transcribed at IE times of infection, IE coding region 2, was unable to induce CAT expression and also did not augment the functional activity of ie1 after cotransfection

A New LCA Methodology of Technology Evolution (TE-LCA) and its Application to the Production of Ammonia (1950-2000) (8 pp)

Goal, Scope and Background: This paper presents a new LCA method of technology evolution (TE-LCA), and its application to the production of ammonia, the second largest chemical product in the world, over the last fifty years. The TE-LCA of a chemical process is the procedure in which historical information on a process, mainly the evolution of technical parameters, is translated by simulation to mass and energy balances as a function of time. These mass and energy balances are then transformed into environmental impact indicators using common LCA approaches. Finally, the evolution of environmental impact resulting from the investigated process can be related to its technical and other, i.e. legislative, developments. Methods: The technological evolution of the production of ammonia was compiled according to three basic sources of information: patents, publications and industry data. From these sources in a first step, the major technological advances of the process were identified as a function of time delivering different process variants that were modelled using the simulation software Aspen Plus®. In a second step, the evolution of environmental regulations is studied. For those energy related emissions that were regulated, e.g. SOx and NOx, it was assumed that threshold values defined in legislation were realized immediately. The aggregation of both steps allows the calculation of the emissions resulting from the production (cradle to gate view) of the investigated chemical as a function of time. Results and Discussion: The application of the TE-LCA to the production of ammonia revealed when and to which extent technological and legislative developments resulted in the reduction of energy related emissions in the production of this chemical compound. Overall, the reduction of emissions from ammonia production was highly influenced by the technological development and only to a lower extent by environmental regulations. Conclusion: The results obtained from the TE-LCA method is useful to reveal how the environmental performance of a process developed in the past and to identify the reasons for this development. The investigated case study of ammonia production shows that investment in technological development also paid off in terms of being ahead of tightened environmental legislation that might bear potential cost consequences such as carbon dioxide tax. Outlook: The presented method can be easily extended by including an economic analysis, which provides additional information on why certain technological developments were enforced and which the economic consequences of changes in environmental legislation were. The new methodology has to be applied to additional case studies, i.e. to other chemical sectors than basic chemicals and to other branches than chemicals. In other chemical sectors, toxic emissions from the production process might have to be considered and trade-offs between these and the overall energy consumption might resul