Molecular Organization of Mason-Pfizer Monkey Virus Capsids Assembled from Gag Polyprotein in Escherichia coli

We describe the results of a study by electron microscopy and image processing of Gag protein shells—immature capsids—of Mason-Pfizer monkey virus assembled in Escherichia coli from two truncated forms of the Gag precursor: Δp4Gag, in which the C-terminal p4Gag was deleted, and Pro(−)CA.NC, in which the N-terminal peptides and proline 1 of the CA domain were deleted. Negative staining of capsids revealed small patches of holes forming a trigonal or hexagonal pattern most clearly visible on occasional tubular forms. The center-to-center spacing of holes in the network was 7.1 nm in Δp4Gag capsids and 7.4 nm in Pro(−)CA.NC capsids. Image processing of Δp4Gag tubes revealed a hexagonal network of holes formed by six subunits with a single subunit shared between rings. This organization suggests that the six subunits are contributed by three trimers of the truncated Gag precursor. Similar molecular organization was observed in negatively stained Pro(−)CA.NC capsids. Shadowed replicas of freeze-etched capsids produced by either construct confirmed the presence of a hexagonal network of holes with a similar center-to-center spacing. We conclude that the basic building block of the cage-like network is a trimer of the Δp4Gag or Pro(−)CA.NC domains. In addition, our results point to a key role of structurally constrained CA domain in the trimeric interaction of the Gag polyprotein

Distinct Roles for Nucleic Acid in In Vitro Assembly of Purified Mason-Pfizer Monkey Virus CANC Proteins

In contrast to other retroviruses, Mason-Pfizer monkey virus (M-PMV) assembles immature capsids in the cytoplasm. We have compared the ability of minimal assembly-competent domains from M-PMV and human immunodeficiency virus type 1 (HIV-1) to assemble in vitro into virus-like particles in the presence and absence of nucleic acids. A fusion protein comprised of the capsid and nucleocapsid domains of Gag (CANC) and its N-terminally modified mutant (ΔProCANC) were used to mimic the assembly of the viral core and immature particles, respectively. In contrast to HIV-1, where CANC assembled efficiently into cylindrical structures, the same domains of M-PMV were assembly incompetent. The addition of RNA or oligonucleotides did not complement this defect. In contrast, the M-PMV ΔProCANC molecule was able to assemble into spherical particles, while that of HIV-1 formed both spheres and cylinders. For M-PMV, the addition of purified RNA increased the efficiency with which ΔProCANC formed spherical particles both in terms of the overall amount and the numbers of completed spheres. The amount of RNA incorporated was determined, and for both rRNA and MS2-RNA, quantities similar to that of genomic RNA were encapsidated. Oligonucleotides also stimulated assembly; however, they were incorporated into ΔProCANC spherical particles in trace amounts that could not serve as a stoichiometric structural component for assembly. Thus, oligonucleotides may, through a transient interaction, induce conformational changes that facilitate assembly, while longer RNAs appear to facilitate the complete assembly of spherical particles

Climate change impact on grape growing in Serbia

Serbian grape growing and fruit production in general is mostly based on relatively small-scale farms with localized terrain and climate features. Climate change analysis and impact studies significantly contribute to the future strategic planning in economic development, and thereby must be done with high level of confidence, which considers cooperative work of the climate research and user community. During the last decade, climate change research in Serbia progressed in accordance with the latest global climate research topics and available computer resources, which are unconditional priority for the quality of the initial data. In this paper are presented latest results of the high-resolution regional climate model simulation, using NMMB model forced with RCP8.5 IPCC scenario (2013). Model bias correction is done for the purpose of their use in impact studies in Serbia. Outcome of the data treatment for model bias reduction is creating a database of the high-resolution daily climate simulation data for the 21st century available for further use in agriculture as well as in other sectors of economy. Usefulness of these data is demonstrated with analysis of the basic climate indices change during the 21st century, which provides assessment of the heat and water availability change, shift in the growing season dates, and risk of the temperature extremes

Climate change impact on grape growing in Serbia

Serbian grape growing and fruit production in general is mostly based on relatively small-scale farms with localized terrain and climate features. Climate change analysis and impact studies significantly contribute to the future strategic planning in economic development, and thereby must be done with high level of confidence, which considers cooperative work of the climate research and user community. During the last decade, climate change research in Serbia progressed in accordance with the latest global climate research topics and available computer resources, which are unconditional priority for the quality of the initial data. In this paper are presented latest results of the high-resolution regional climate model simulation, using NMMB model forced with RCP8.5 IPCC scenario (2013). Model bias correction is done for the purpose of their use in impact studies in Serbia. Outcome of the data treatment for model bias reduction is creating a database of the high-resolution daily climate simulation data for the 21st century available for further use in agriculture as well as in other sectors of economy. Usefulness of these data is demonstrated with analysis of the basic climate indices change during the 21st century, which provides assessment of the heat and water availability change, shift in the growing season dates, and risk of the temperature extremes

Fine scale climate change analysis: From global models to local impact studies in serbia

Climate change studies involve changes in climate system of global long-term scales with effects on regional and local climates, down to shortrange time scales, like weather hazards and extremes. Climate studies significantly contribute to the future strategic planning in economic development and thereby must produce results with high level of confidence. Global climate models simulations of the past and future climate are initial step in climate change research, but their coarse resolution data are unable to provide useful information for analysis on national levels. In this paper is presented workflow algorithm of the procedures necessary to convert coarse global climate system projected changes to fine scale data with included effect of small scale features and reduced model bias impact

Climate change impact on grape growing in Serbia

Serbian grape growing and fruit production in general is mostly based on relatively small-scale farms with localized terrain and climate features. Climate change analysis and impact studies significantly contribute to the future strategic planning in economic development, and thereby must be done with high level of confidence, which considers cooperative work of the climate research and user community. During the last decade, climate change research in Serbia progressed in accordance with the latest global climate research topics and available computer resources, which are unconditional priority for the quality of the initial data. In this paper are presented latest results of the high-resolution regional climate model simulation, using NMMB model forced with RCP8.5 IPCC scenario (2013). Model bias correction is done for the purpose of their use in impact studies in Serbia. Outcome of the data treatment for model bias reduction is creating a database of the high-resolution daily climate simulation data for the 21st century available for further use in agriculture as well as in other sectors of economy. Usefulness of these data is demonstrated with analysis of the basic climate indices change during the 21st century, which provides assessment of the heat and water availability change, shift in the growing season dates, and risk of the temperature extremes

PTEN Sequence Analysis in Endometrial Hyperplasia and Endometrial Carcinoma in Slovak Women

Phosphatase and tensin homolog (PTEN) is a protein that acts as a tumor suppressor by dephosphorylating the lipid second messenger phosphatidylinositol 3,4,5-trisphosphate. Loss of PTEN function has been implicated in the pathogenesis of a number of different tumors, particularly endometrial carcinoma (ECa). ECa is the most common neoplasia of the female genital tract. Our study evaluates an association between the morphological appearance of endometrial hyperplasia and endometrial carcinoma and the degree of PTEN alterations. A total of 45 endometrial biopsies from Slovak women were included in present study. Formalin-fixed and paraffin-embedded tissue samples with simple hyperplasia (3), complex hyperplasia (5), atypical complex hyperplasia (7), endometrioid carcinomas G1 (20) and G3 (5), and serous carcinoma (5) were evaluated for the presence of mutations in coding regions of PTEN gene, the most frequently mutated tumor suppressor gene in endometrial carcinoma. 75% of the detected mutations were clustered in exons 5 and 8. Out of the 39 mutations detected in 24 cases, 20 were frameshifts and 19 were nonsense, missense, or silent mutations. Some specimens harboured more than one mutation. The results of current study on Slovak women were compared to a previous study performed on Polish population. The two sets of results were similar