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tRNA-like structures of plant viral RNAs: conformational requirements for adenylation and aminoacylation.

Bromo- and cucumovirus RNAs contain a tRNA-like structure as an integral part of their genome. This structure is located at the 3' end of the viral RNA and is an acceptor of tyrosine. The 3' regions of representative viral RNAs have been sequenced and quite unorthodox secondary foldings have been proposed for these 3' ends. The question therefore remained as to how these structures could be recognized by tRNA-specific enzymes. We have established the minimum number of nucleotides from the 3' end of the brome mosaic virus and broad bean mottle virus RNAs required for the formation of structures recognized by the tyrosyl-tRNA synthetase and/or the tRNA nucleotidyltransferase. The results obtained delineate the length of the tRNA-like region, and indicate that the 5' region of the tRNA-like structure participates in the formation of the amino acid stem. This has led us to propose an 'L'-shaped secondary structure for these tRNA-like regions.

Anticodon↗

Crystal structure of plant asparaginase.

In plants, specialized enzymes are required to catalyze the release of ammonia from asparagine, which is the main nitrogen-relocation molecule in these organisms. In addition, K+-independent plant asparaginases are also active in splitting the aberrant isoaspartyl peptide bonds, which makes these proteins important for seed viability and germination. Here, we present the crystal structure of potassium-independent L-asparaginase from yellow lupine (LlA) and confirm the classification of this group of enzymes in the family of Ntn-hydrolases. The alpha- and beta-subunits that form the mature (alphabeta)2 enzyme arise from autoproteolytic cleavage of two copies of a precursor protein. In common with other Ntn-hydrolases, the (alphabeta) heterodimer has a sandwich-like fold with two beta-sheets flanked by two layers of alpha-helices (alphabetabetaalpha). The nucleophilic Thr193 residue, which is liberated in the autocatalytic event at the N terminus of subunit beta, is part of an active site that is similar to that observed in a homologous bacterial enzyme. An unusual sodium-binding loop of the bacterial protein, necessary for proper positioning of all components of the active site, shows strictly conserved conformation and metal coordination in the plant enzyme. A chloride anion complexed in the LlA structure marks the position of the alpha-carboxylate group of the L-aspartyl substrate/product moiety. Detailed analysis of the active site suggests why the plant enzyme hydrolyzes asparagine and its beta-peptides but is inactive towards substrates accepted by similar Ntn-hydrolases, such as taspase1, an enzyme implicated in some human leukemias. Structural comparisons of LlA and taspase1 provide interesting insights into the role of small inorganic ions in the latter enzyme.

Amidohydrolases↗

The Structure of Plant Cell Walls: II. The Hemicellulose of the Walls of Suspension-cultured Sycamore Cells.

The molecular structure, chemical properties, and biological function of the xyloglucan polysaccharide isolated from cell walls of suspension-cultured sycamore (Acer pseudoplatanus) cells are described. The sycamore wall xyloglucan is compared to the extracellular xyloglucan secreted by suspension-cultured sycamore cells into their culture medium and is also compared to the seed "amyloid" xyloglucans.Xyloglucan-or fragments of xyloglucan-and acidic fragments of the pectic polysaccharides are released from endopolygalacturonase-pretreated sycamore walls by treatment of these walls with 8 m urea, endoglucanase, or 0.5 n NaOH. Some of the xyloglucan thus released is found to cochromatograph with the acidic pectic fragments on diethylaminoethyl Sephadex. The chemical or enzymic treatments required for the release of xyloglucan from the walls and the cochromatography of xyloglucan with the acidic pectic fragments indicate that xyloglucan is covalently linked to the pectic polysaccharides and is noncovalently bound to the cellulose fibrils of the sycamore cell wall.The molecular structure of sycamore xyloglucan was characterized by methylation analysis of the oligosaccharides obtained by endoglucanase treatment of the polymer. The structure of the polymer is based on a repeating heptasaccharide unit which consists of 4 residues of beta-1-4-linked glucose and 3 residues of terminal xylose. A single xylose residue is glycosidically linked to carbon 6 of 3 of the glucosyl residues.

Journal Article↗

The movement proteins of cowpea mosaic virus and cauliflower mosaic virus induce tubular structures in plant and insect cells.

The movement proteins (MP) of cowpea mosaic virus and cauliflower mosaic virus (CaMV) are associated with tubular structures in vivo which participate in the transmission of virus particles from cell to cell. Both proteins have been expressed in plant protoplasts and insect cells. In all cases, immunofluorescent histochemistry showed that the MPs accumulate intracellularly as tubular extensions projecting from the cell surface. Additionally, electron microscopy revealed intracellular MP aggregates in CaMV MP-expressing cells. The data presented establish common features for the tubule-forming MPs: no other virus gene products are required for tubule formation and unique plant components (e.g. plasmodesmata) are not essential for tubule synthesis.

Animals↗

Possible secondary structure in plant and yeast beta-glucanase.

Circular-dichroism spectra of a barley 1,3-beta-glucanase were analysed by two methods. The combined results predict 36-40% helix and 15-18% beta-structure in the protein. Prediction of secondary-structural features on the basis of amino acid sequence information yielded overall helix and beta-structure contents of 37% and 19% respectively. Comparison of the predicted structural elements along the barley 1,3-beta-glucanase with those of three related plant glucanases and a yeast glucanase suggest a close similarity in secondary structure among the five proteins. Consideration is given to the potential importance of certain amino acids which are conserved in these five glucanases.

Amino Acid Sequence↗

Structures of plant viruses from vibrational circular dichroism.

Vibrational circular dichroism (VCD) spectra in the amide I and II regions have been measured for viruses for the first time. VCD spectra were recorded for films prepared from aqueous buffer solutions and also for solutions using D(2)O buffers at pH 8. Investigations of four filamentous plant viruses, Tobacco mosaic virus (TMV), Papaya mosaic virus, Narcissus mosaic virus (NMV) and Potato virus X (PVX), as well as a deletion mutant of PVX, are described in this paper. The film VCD spectra of the viruses clearly revealed helical structures in the virus coat proteins; the nucleic acid bases present in the single-stranded RNA could also be characterized. In contrast, the solution VCD spectra showed the characteristic VCD bands for alpha-helical structures in the coat proteins but not for RNA. Both sets of results clearly indicated that the coat protein conformations are dominated by helical structures, in agreement with earlier reports. VCD results also indicated that the coat protein structures in PVX and NMV are similar to each other and somewhat different from that of TMV. The present study demonstrates the feasibility of measuring VCD spectra for viruses and extracting structural information from these spectra.

Capsid Proteins↗

The eight-cysteine motif, a versatile structure in plant proteins.

A number of protein sequences deduced from the molecular analysis of plant cDNA or genomic libraries can be grouped in relation to a defined number of cysteine residues located in distinct positions of their sequences. This is the case for a group of around 500 polypeptides from different species that contain a small domain (less than 100 amino acids residues) displaying a pattern of eight-cysteines in a specific order. The plant sequences containing this motif belong to proteins having different functions, ranging from storage, protection, enzyme inhibition and lipid transfer, to cell wall structure. The eight-cysteine motif (8CM) appears to be a structural scaffold of conserved helical regions connected by variable loops, as observed by three-dimensional structure analysis. It is proposed that the cysteine residues would form a network of disulfide bridges necessary, for the maintenance of the tertiary structure of the molecule together with the central helical core, while the variable loops would provide the sequences required for the specific functions of the proteins.

Amino Acid Motifs↗

The crystal structure of plant ATG12 and its biological implication in autophagy.

Atg12 is a post-translational modifier that is activated and conjugated to its single target, Atg5, by a ubiquitin-like conjugation system. The Atg12-Atg5 conjugate is essential for autophagy, the bulk degradation process of cytoplasmic components by the vacuolar/lysosomal system. Here, we demonstrate that the Atg12 conjugation system exists in Arabidopsis and is essential for plant autophagy as well as in yeast and mammals. We also report the crystal structure of Arabidopsis thaliana (At) ATG12 at 1.8 A resolution. Despite no obvious sequence homology with ubiquitin, the structure of AtATG12 shows a ubiquitin fold strikingly similar to those of mammalian homologs of Atg8, the other ubiquitin-like modifier essential for autophagy, which is conjugated to phosphatidylethanolamine. Two types of hydrophobic patches are present on the surface of AtATG12: one is conserved in both Atg12 and Atg8 orthologs, while the other is unique to Atg12 orthologs. Considering that they share Atg7 as an E1-like enzyme, we suggest that the first hydrophobic patch is responsible for the conjugation reaction, while the latter is involved in Atg12-specific functions.

Amino Acid Sequence↗

Prediction of three-dimensional structure of plant lectins from the domains of concanavalin A.

The circularly permuted sequence homology that relates concanavalin A to the other leguminous plant lectins can be explained by an evolutionary model that requires three exons. The identification of these potential exons from the amino acid sequence data allows the prediction of three domains in the lectin structure. The predicted domains are reasonable, functional and structural units in concanavalin A, demonstrating the correspondence of exons and domains. In addition, slight modifications to the three-dimensional structure of concanavalin A produced a model which was consistent with the sequence data for the other plant lectins.

Amino Acid Sequence↗

RESEARCH: Plant Community Structure in Relation to Long-Term Disturbance by Mechanized Military Maneuvers in a Semiarid Region.

/ Mechanized military maneuvers are an intensive form of disturbance to plant communities in large areas throughout the world. Tracking by heavy vehicles can cause direct mortality and indirectly affect plant communities through soil compaction and by altering competitive relationships. We assessed the long-term condition of structural attributes of open woodland, grassland, and shrubland communities at Fort Carson, Colorado, in relation to levels of disturbance and soil texture. Covariate analyses were used to help separate the directional forcings by the chronic disturbance from the regenerative capacities in order to assess the relative resistance and resilience of the communities and to determine whether the continual disturbance-recovery processes balanced under current levels of utilization. All three communities responded differently to disturbance. In open woodlands, altered understory/overstory relationships were suggested by increased grass, forb, shrub, and total vegetation cover and smaller decreases in shorter than taller woody species with increasing levels of disturbance. Grassland communities generally displayed greater responses to disturbance than other communities, but temporal dynamics were often similar, indicating relatively less resistance but greater resilience of this community. Weed and exotic species increased both temporally and in relation to levels of disturbance in all three community types. Temporal trends in community-level indices of dissimilarity and diversity also indicate that rates of disturbance were greater than rates of recovery. Few variables were related to within-community differences in soil texture. While total aerial cover was temporally stable, changes in species composition and in basal cover in grasslands and shrublands suggest increasing erosion potential.

Journal Article↗

A dynamic growth model of vegetative soya bean plants: model structure and behaviour under varying root temperature and nitrogen concentration.

A differential equation model of vegetative growth of the soya bean plant (Glycine max (L.) Merrill cv. Ransom') was developed to account for plant growth in a phytotron system under variation of root temperature and nitrogen concentration in nutrient solution. The model was tested by comparing model outputs with data from four different experiments. Model predictions agreed fairly well with measured plant performance over a wide range of root temperatures and over a range of nitrogen concentrations in nutrient solution between 0.5 and 10.0 mmol NO3- in the phytotron environment. Sensitivity analyses revealed that the model was most sensitive to changes in parameters relating to carbohydrate concentration in the plant and nitrogen uptake rate.

Carbohydrate Metabolism↗

Tumor DNA structure in plant cells transformed by A. tumefaciens.

Crown gall tumors are induced in plants by infection with the soil bacterium Agrobacterium tumefaciens. Because the tumor induction involves transfer of a portion of the tumor-inducing (Ti) plasmid DNA from the bacterium to the plant cells, this system is of interest for the study of genetic exchange as well as tumor induction. The boundaries of the transferred DNA (T-DNA) have been cloned from transformed plant cells of tobacco. Detailed mapping with restriction enzymes and nucleotide sequence analysis of two independent clones were used to study the molecular structure of the ends of the T-DNA. One clone contains the two ends of the T-DNA joined together; the other contains one end of the T-DNA joined to repetitive plant DNA sequences. These studies provide direct evidence that the T-DNA can be integrated into the plant genome. In addition, the data suggest that in the plant, T-DNA can be tandemly repeated. Sequence analysis of the junction of crown gall clone 1 reveals several direct repeats as well as an inverted repeat; these structures may be involved in the transfer of the DNA from Agrobacterium to plant cells.

Base Sequence↗

Leading structures of plant origin for drug development.

This review summarizes the most important results of a plant screening program performed in the author's laboratory during the last 10 years. The screening program was initiated to search for new plant constituents with potential pharmacological activity in the field of inflammation in general, allergic asthma, immunodeficiencies and cardiovascular diseases. The test program mainly utilised in vitro assays, and particular attention was given to the key enzymes of the prostaglandin metabolism, cellular and humoral targets of the unspecific immune system, an in vitro ACE-inhibition test system, and an in vivo plethysmographic system as an inhibitory model of bronchial obstruction. The results of structure-activity relationship studies are also described.

Animals↗

Effects of weightlessness on photosynthesizing cells structure of plant.

It was established early that the signs of accelerated aging of both plant leaves and of moss chloronema were observed in seedlings of high plants and in Funaria hygrometrica protonema under long term growing under weightlessness. It was observed the structure changing of photosynthesizing cells in Arabidopsis thaliana, Epidendrum radicans and Pisum sativum leaves. Authors found out the extension and vesiculation of thylakoids between chloroplast granas. They observed thylakoids partly destroying under 96 and 110 days microgravity influence. But some questions are still opened: 1. Are these changes consequence of accelerated differentiation and aging photosynthesizing cells? 2. Do the definite changes appear in photosynthesizing cells during short-term microgravity influence? Therefore the study of mesophyll cells ultrastructure of leaves that finished growth by tension in microgravity was the idea of our experiment.

Adenosine Triphosphate↗

The plant dehydrins: structure and putative functions.

This review deals with recent data on the structure and biochemical properties of dehydrins, proteins that are normally synthesized in maturating seeds during their desiccation, and also in vegetative tissues of plants treated with abscisic acid or exposed to environmental stress factors that result in cellular dehydration. The dehydrins are considered as stress proteins involved in formation of plant protective reactions against dehydration. The generally accepted classification of dehydrins is based on their structural features, such as the presence of conserved sequences, designated as Y-, S-, and K-segments. The K-segment representing a highly conserved 15 amino acid motif (EKKGIMDKIKEKLPG) forming amphiphilic alpha-helix has been found in all dehydrins. The pathways of regulation of dehydrin gene expression, putative functions of dehydrins, and molecular mechanisms of their actions are discussed.

Gene Expression Regulation, Plant↗