Etude des modifications du système endomembranaire lors de la réplication du Grapevine fanleaf virus

STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

3D structure prediction of histone acetyltransferase (HAC) proteins of the p300/CBP family and their interactome in Arabidopsis thaliana

Histone acetylation is an important posttranslational modification correlated with gene activation. In Arabidopsis thaliana the histone acetyltransferase (HAC) proteins of the CBP family are homologous to animal p300/CREB (cAMP-responsive element-binding proteins, which are important histone acetyltransferases participating in many physiological processes, including proliferation, differentiation, and apoptosis. In this study the 3-D structure of all HAC protein subunits in Arabidopsis thaliana: HAC1, HAC2, HAC4, HAC5 and HAC12 is predicted by homology modeling and confirmed by Ramachandran plot analysis. The amino acid sequences HAC family members are highly similar to the sequences of the homologous human p300/CREB protein. Conservation of p300/CBP domains among the HAC proteins was examined further by sequence alignment and pattern search. The domains of p300/CBP required for the HAC function, such as PHD, TAZ and ZZ domains, are conserved in all HAC proteins. Interactome analysis revealed that HAC1, HAC5 and HAC12 proteins interact with S-adenosylmethionine-dependent methyltransferase domaincontaining protein that shows methyltransferase activity, suggesting an additional function of the HAC proteins. Additionally, HAC5 has a strong interaction value for the putative c-myb-like transcription factor MYB3R-4, which suggests that it also may have a function in regulation of DNA replication

Challenging the role of microtubules in Tobacco mosaic virus movement by drug treatments is disputable

The movement protein (MP) of Tobacco mosaic virus interacts with microtubules during infection. Although this interaction is correlated with the function of MP in the cell-to-cell transport of viral RNA, a direct role of microtubules in the movement process was recently challenged by studies involving the treatment of plants with inhibitors of microtubule polymerization. Here, we report evidence suggesting that such treatments may not efficiently disrupt all microtubules. Thus, results obtained from studies using microtubule inhibitors may have to remain open to interpretation with regard to the involvement of microtubules in viral RNA trafficking

Biosynthesis and characterization of silver nanoparticles using Trichoderma longibrachiatum and their effect on phytopathogenic fungi

Abstract An efficient biosynthesis process for the rapid production of nanoparticles would enable the development of a “microbial nanotechnology” for mass-scale production. In the present research, biological silver nanoparticle was synthesized extracellularly by using the fungus, Trichoderma longibrachiatum, where the cell filtrate of the fungus was used as a reducing and stabilizing agent in the process of nanoparticle synthesis. Different physical parameters such as fungal biomass concentration (1, 5, 10, 15, and 20 g), temperature (25, 28, and 33 °C), incubation time (0–120 h), and agitation (shaken or not shaken) were investigated, in order to determine the optimal conditions for nanoparticle biosynthesis. The stability and antifungal properties of the synthesized silver nanoparticles (AgNPs) were also determined. Data revealed that a combination of 10 g fungal biomass, a reaction temperature of 28 °C, a 72-h incubation time, and without shaking were the optimum conditions for the synthesis of the silver nanoparticles. Visual observation of brown color is an indication of silver nanoparticle production. UV–vis spectroscopy showed maximum absorption at 385 nm with the optimum conditions. Transmission electron microscopy (TEM) revealed the formation of monodispersed spherical shape with a mean diameter of 10 nm. Fourier transformation infrared (FTIR) showed bands at1634.92 and 3269.31 cm−1. Dynamic light scattering (DLS) supported that the Z-average size was 24.43 and 0.420 PdI value. Zeta potential showed − 19.7 mV with a single peak. The AgNPs synthesized through this biosystem approach were relatively stable up to 2 months after synthesis. The use of AgNPs as antifungal led to significant reductions in the number of forming colonies for many plant pathogenic fungi, with efficiencies reaching up to 90% against Fusarium verticillioides, Fusarium moniliforme, Penicillium brevicompactum, Helminthosporium oryzae, and Pyricularia grisea. However, further research should be carried out in order to determine the toxic effect of AgNPs before mass production and use of agricultural applications

Dynamics of COPII Vesicles and the Golgi Apparatus in Cultured Nicotiana tabacum BY-2 Cells Provides Evidence for Transient Association of Golgi Stacks with Endoplasmic Reticulum Exit Sites

Despite the ubiquitous presence of the COPI, COPII, and clathrin vesicle budding machineries in all eukaryotes, the organization of the secretory pathway in plants differs significantly from that in yeast and mammalian cells. Mobile Golgi stacks and the lack of both transitional endoplasmic reticulum (ER) and a distinct ER-to-Golgi intermediate compartment are the most prominent distinguishing morphological features of the early secretory pathway in plants. Although the formation of COPI vesicles at periphery of Golgi cisternae has been demonstrated in plants, exit from the ER has been difficult to visualize, and the spatial relationship of this event is now a matter of controversy. Using tobacco (Nicotiana tabacum) BY-2 cells, which represent a highly active secretory system, we have used two approaches to investigate the location and dynamics of COPII binding to the ER and the relationship of these ER exit sites (ERES) to the Golgi apparatus. On the one hand, we have identified endogenous COPII using affinity purified antisera generated against selected COPII-coat proteins (Sar1, Sec13, and Sec23); on the other hand, we have prepared a BY-2 cell line expressing Sec13:green fluorescent protein (GFP) to perform live cell imaging with red fluorescent protein–labeled ER or Golgi stacks. COPII binding to the ER in BY-2 cells is visualized as fluorescent punctate structures uniformly distributed over the surface of the ER, both after antibody staining as well as by Sec13:GFP expression. These structures are smaller and greatly outnumber the Golgi stacks. They are stationary, but have an extremely short half-life (<10 s). Without correlative imaging data on the export of membrane or lumenal ER cargo it was not possible to equate unequivocally these COPII binding loci with ERES. When a GDP-fixed Sar1 mutant is expressed, ER export is blocked and the visualization of COPII binding is perturbed. On the other hand, when secretion is inhibited by brefeldin A, COPII binding sites on the ER remain visible even after the Golgi apparatus has been lost. Live cell imaging in a confocal laser scanning microscope equipped with spinning disk optics allowed us to investigate the relationship between mobile Golgi stacks and COPII binding sites. As they move, Golgi stacks temporarily associated with COPII binding sites at their rims. Golgi stacks were visualized with their peripheries partially or fully occupied with COPII. In the latter case, Golgi stacks had the appearance of a COPII halo. Slow moving Golgi stacks tended to have more peripheral COPII than faster moving ones. However, some stationary Golgi stacks entirely lacking COPII were also observed. Our results indicate that, in a cell type with highly mobile Golgi stacks like tobacco BY-2, the Golgi apparatus is not continually linked to a single ERES. By contrast, Golgi stacks associate intermittently and sometimes concurrently with several ERES as they move