Tyrosine Phosphorylation of Caspase-8 Abrogates Its Apoptotic Activity and Promotes Activation of c-Src

Src family tyrosine kinases (SFKs) phosphorylate caspase-8A at tyrosine (Y) 397 resulting in suppression of apoptosis. In addition, the phosphorylation of caspase-8A at other sites including Y465 has been implicated in the regulation of caspase-8 activity. However, the functional consequences of these modifications on caspase-8 processing/activity have not been elucidated. Moreover, various Src substrates are known to act as potent Src regulators, but no such role has been explored for caspase-8. We asked whether the newly identified caspase-8 phosphorylation sites might regulate caspase-8 activation and conversely, whether caspase-8 phosphorylation might affect Src activity. Here we show that Src phosphorylates caspase-8A at multiple tyrosine sites; of these, we have focused on Y397 within the linker region and Y465 within the p12 subunit of caspase-8A. We show that phosphomimetic mutation of caspase-8A at Y465 prevents its cleavage and the subsequent activation of caspase-3 and suppresses apoptosis. Furthermore, simultaneous phosphomimetic mutation of caspase-8A at Y397 and Y465 promotes the phosphorylation of c-Src at Y416 and increases c-Src activity. Finally, we demonstrate that caspase-8 activity prevents its own tyrosine phosphorylation by Src. Together these data reveal that dual phosphorylation converts caspase-8 from a pro-apoptotic to a pro-survival mediator. Specifically, tyrosine phosphorylation by Src renders caspase-8 uncleavable and thereby inactive, and at the same time converts it to a Src activator. This novel dynamic interplay between Src and caspase-8 likely acts as a potent signal-integrating switch directing the cell towards apoptosis or survival.     

Temporal variability in the spatial and environmental determinants of functional metacommunity organization – stream fish in a human‐modified landscape

1. Quantifying the relative importance of environmental filtering versus regional spatial structuring has become an intensively studied area in the context of metacommunity ecology. However, most studies have evaluated the role of environmental and spatial processes using taxonomic data sets of single snapshot surveys. 2. Here, we examined temporal changes in patterns and possible processes behind the functional metacommunity organization of stream fishes in a human‐modified landscape. Specifically, we (i) studied general changes in the functional composition of fish assemblages among 40 wadeable stream sites during a 3‐year study period in the catchment area of Lake Balaton, Hungary, (ii) quantified the relative importance of spatial and environmental factors as determinants of metacommunity structure and (iii) examined temporal variability in the relative role of spatial and environmental processes for this metacommunity. 3. Partial triadic analysis showed that assemblages could be effectively ordered along a functional gradient from invertebrate consuming species dominated by the opportunistic life‐history strategy, to assemblages with a diverse array of functional attributes. The analysis also revealed that functional fish assemblage structure was moderately stable among the sites between the sampling periods. 4. Despite moderate stability, variance partitioning using redundancy analyses (RDA) showed considerable temporal variability in the contribution of environmental and spatial factors to this pattern. The analyses also showed that environmental variables were, in general, more important than spatial ones in determining metacommunity structure. Of these, natural environmental variables (e.g. altitude, velocity) proved to be more influential than human‐related effects (e.g. pond area, % inhabited area above the site, nutrient enrichment), even in this landscape with relatively low variation in altitude and stream size. 5. Pond area was, however, the most important human stressor variable that was positively associated with the abundance of non‐native species with diverse functional attributes. The temporal variability in the relative importance of environmental and spatial factors was probably shaped by the release of non‐native fish from fish ponds to the stream system during flood events. 6. To conclude, both spatial processes and environmental control shape the functional metacommunity organization of stream fish assemblages in human‐modified landscapes, but their importance can vary in time. We argue, therefore, that metacommunity studies should better consider temporal variability in the ecological mechanisms (e.g. dispersal limitation, species sorting) that determine the dynamics of landscape‐level community organization.     

Biosynthesis of chlorophyll P-680 of photosystem II in greening leaves of plants adapted to heat shock

In etiolated pea and maize leaves illuminated after incubation at 38 degreesC, a new dark reaction was shown manifested in the bathochromic shift of spectral bands and accompanied by esterification of the product of protochlorophyllide photochemical reduction--Chld 684/676: Chld 684/676 --> Chl 688/680. After completion of the reaction a rapid (20-30 sec) quenching of the fluorescence of the reaction product (Chl 688/680) was observed. The reaction Chld 684/676 --> Chl 688/680 is inhibited under anaerobic conditions and in the presence of cyanide; the reaction accompanied by Chl 688/680 fluorescence quenching is not observed in pea mutants with impaired function of photosystem II reaction centers. The spectral properties of the formed Chl form with the absorption maximum at 680 nm, fluorescence quenching, and simultaneous synthesis of pheophytin suggest that the reaction is connected with the chlorophyll of photosystem II reaction center--P-680.     

Effect of nitrogen nutrition on Fusarium wilt of tomato plants

The role of nitrate-nitrogen (NO₃-N) in relation to the development of tomato wilt caused by Fusarium oxysporum f. sp. lycopersici R1 was studied. Plants receiving 284 μg/ml nitrogen in the nutrient solution exhibited the same severe wilt symptoms as the control plants in soil. Disease decreased with increasing nitrogen levels (420, 630 and 1050 μg/ml). Apparently, the plants were also less susceptible to the disease when the concentration of nitrogen was 70 μg/ml, i. e. below the optimal level (284 μg/ml). Protein content in tissues of plants grown with different amounts of nitrogen was also determined. High nitrogen levels, which decreased disease severity, increased the protein content in leaf tissues. Of 17 amino acids only proline content increased with increasing nitrogen supply. High doses of NO₃–N decreased the phenol content and the activity of peroxidase in stem and leaf tissues of tomato plants. It is suggested that phenolic compounds and the activity of peroxidase are not significant in the resistance of tomato to Fusarlum wilt associated with high nitrogen supply.     

Rainforest and savanna landscape dynamics in New Caledonia: Towards a mosaic of stable rainforest and savanna states?

Stable forested environments can be converted to savanna in response to changes in environmental disturbances. New Caledonia is a biodiversity hotspot; significant ecological and economic resources would be lost if forests were turned into savanna by anthropogenic environmental changes. On the landscape scale, systems that have undergone shifts of this kind are characterized by sharp forest–savanna boundaries and mosaic‐like distributions of savanna and forest. Understanding the locations and the dynamics of such boundaries is a challenge for ecologists and is critical for landscape management and biodiversity conservation. Using a time series of aerial photographs (1955–2000) and a forest habitat suitability map, we tested the hypothesis that topography and spatial processes, especially those relating to fire spread and seed dispersal, are the main determinants of the spatial distribution of rainforest and savanna in a New Caledonian landscape covering 24 km². Within the studied landscape, the overall forest coverage decreased by 24% between 1976 and 2000. This was primarily due to the contraction of forests on west‐facing slopes, which accounted for about 90% of the total loss. Conversely, the east‐facing forests seemed to have contracted extensively prior to the studied period, and were confined to refuges. A habitat suitability index calculated from the landscape's topographical features using generalized additive models accurately predicted both the presence of forests and the probability of forest expansion/contraction. We also provide evidence that spatial processes such as fire spread and seed dispersal limit the expansion and contraction of forests. Our results suggest that rainforests on west‐facing slopes in New Caledonia will be progressively destroyed by fire until they are restricted to refuges along thalwegs and creeks, as appears to have already happened for their east‐facing counterparts.