Membrane translocation and oligomerization of hBok are triggered in response to apoptotic stimuli and Bnip3

Abstract.: hBok is a human pro-apoptotic member of the Bcl-2 family. By fluorescence in situ hybridization and in silico analysis, hBok was found to be located on chromosome 2q37.3. Its expression was detected in various organs and several hormonally regulated cancer cells. Expression of hBok was shown to be upregulated in estrogen-dependent breast cancer by estrogen deprivation and in myocardial cells during hypoxia. Confocal laser scanning microscopy examinations and subcellular fractionation studies showed that hBok was distributed in both the cytosol and intracellular membranes of healthy cells. Upon overexpression of hBok or stimulation of apoptosis, hBok became integrated into the membrane. Furthermore, apoptosis and oligomerization were promoted by BH3-only proteins, such as Bid, Bnip3 and p53, but prevented by BFL-1. hBok was found to interact with Bnip3. Our findings suggest that functional BH3-only proteins facilite the oligomerization and insertion of hBok into the membrane to activate i

Macro- and microscopic leaf characteristics of six grapevine genotypes (Vitis spp.) with different susceptibilities to grapevine downy mildew

This work reports the leaf morphology of six grapevine genotypes, five belonging to Vitis vinifera and one to Vitis riparia. Earlier studies on these genotypes showed different levels of susceptibility to grapevine downy mildew (Plasmopara viticola). The aim of this work was to detect differences between the leaf morphology of these cultivars at the macro- and microscopic levels, and to characterize morphological traits which could be associated with susceptibility and resistance to downy mildew. An ampelographic description of each genotype was used to develop a scheme illustrating the characteristic leaf morphology. The density and morphology of the trichomes and the stomatal index was assessed by means of microscopical techniques. Distinct macro and microscope differences among the genotypes were seen. No clear relation between ampelographic characteristics and susceptibility to downy mildew was observed. The two cultivars that in earlier studies were found to be the least susceptible to downy mildew were the most similar in terms of their spongy mesophyll. Both showed very little or no wax on the abaxial surface of their leaves.

Ancestral chemotypes of cultivated grapevine with resistance to Botryosphaeriaceae-related dieback allocate metabolism towards bioactive stilbenes

- Grapevine trunk diseases have devastating consequences on vineyards worldwide. European wild grapevines (Vitis vinifera subs. sylvestris) from the last viable population in Germany along the Rhine river showed variable degrees of resistance against Neofusicoccum parvum (strain Bt‐67), a fungus associated with Botryosphaeriaceae‐related dieback. - Representative genotypes from different subclades of this population were mapped with respect to their ability to induce wood necrosis, as well as their defence responses in a controlled inoculation system. - The difference in colonization patterns could be confirmed by cryo‐scanning electron microscopy, while there was no relationship between vessel diameter and infection success. Resistant lines accumulated more stilbenes, that were in addition significantly partitioned to nonglycosylated viniferin trimers. By contrast, the susceptible genotypes accumulated less stilbenes with a significantly higher proportion of glycosylated piceid. - We suggest a model in which in the resistant genotypes phenylpropanoid metabolism is channelled rapidly and specifically to the bioactive stilbenes. Our study specifies a resistant chemotype against grapevines trunk diseases and paves a way to breed for resistance against grapevine Botryosphaeriaceae‐related dieback

The mode of interaction between Vitis and Plasmopara viticola Berk. & Curt. Ex de Bary depends on the host species

In order to obtain insight into host responses to grapevine downy mildew (Plasmopara viticola), we compared pathogen development on a panel of Vitis species from North America, Asia and Europe. Leaf discs from different host species were inoculated in parallel, and the colonisation of the mesophyll was visualised by aniline blue staining and quantified with respect to infection incidence and mycelial growth. In parallel, the morphology of guard cells was screened for the presence of an internal cuticular rim after staining with acridine orange and using low-temperature scanning electron microscopy. We observed three response patterns: (i) inhibition of pathogen development early after attachment of zoospores; (ii) successful colonisation of the mesophyll by the pathogen; and (iii) aberrant development, where the pathogen does not attach to guard cells, but produces hyphae on the leaf surface without formation of viable sporangiophores. Inhibition is observed in the North American and Siberian species, successful colonisation prevails in the European hosts, and surface hyphae are found on non-Siberian Asiatic species. We propose that the interaction between host and pathogen is under control of specific signals that have been subject to evolutionary diversification

Neutralizing the Detrimental Effect of Glutathione on Precious Metal Catalysts

We report our efforts to enable transition-metal catalysis in the presence of cellular debris, notably Escherichia coli cell free extracts and cell lysates. This challenging goal is hampered by the presence of thiols, mainly present in the form of glutathione (GSH), which poison precious metal catalysts. To overcome this, we evaluated a selection of oxidizing agents and electrophiles toward their potential to neutralize the detrimental effect of GSH on a Ir-based transfer hydrogenation catalyst. While the bare catalyst was severely inhibited by cellular debris, embedding the organometallic moiety within a host protein led to promising results in the presence of some neutralizing agents. In view of its complementary to natural enzymes, the asymmetric imine reductase based on the incorporation of a biotinylated iridium pianostool complex within streptavidin (Sav) isoforms was selected as a model reaction. Compared to purified protein samples, we show that pretreatment of cell free extracts and cell lysates containing Sav mutants with diamide affords up to <100 TON’s and only a modest erosion of enantioselectivity

Macro- and microscopic leaf characteristics of six grapevine genotypes (Vitis spp.) with different susceptibilities to grapevine downy mildew

This work reports the leaf morphology of six grapevine genotypes, five belonging to Vitis vinifera and one to Vitis riparia. Earlier studies on these genotypes showed different levels of susceptibility to grapevine downy mildew (Plasmopara viticola). The aim of this work was to detect differences between the leaf morphology of these cultivars at the macro- and microscopic levels, and to characterize morphological traits which could be associated with susceptibility and resistance to downy mildew. An ampelographic description of each genotype was used to develop a scheme illustrating the characteristic leaf morphology. The density and morphology of the trichomes and the stomatal index was assessed by means of microscopical techniques. Distinct macro and microscope differences among the genotypes were seen. No clear relation between ampelographic characteristics and susceptibility to downy mildew was observed. The two cultivars that in earlier studies were found to be the least susceptible to downy mildew were the most similar in terms of their spongy mesophyll. Both showed very little or no wax on the abaxial surface of their leaves.This research was supported by a fellowship from the Fundación Martin Escudero and Ministerio de Educación y Ciencia (Spain).Peer reviewe