Hsp90 chaperone code and the tumor suppressor VHL cooperatively regulate the mitotic checkpoint

Heat shock protein-90 (Hsp90) is an essential molecular chaperone in eukaryotes that plays a vital role in protecting and maintaining the functional integrity of deregulated signaling proteins in tumors. We have previously reported that the stability and activity of the mitotic checkpoint kinase Mps1 depend on Hsp90. In turn, Mps1-mediated phosphorylation Hsp90 regulates its chaperone function and is essential for the mitotic arrest. Cdc14-assisted dephosphorylation of Hsp90 is vital for the mitotic exit. Post-translational regulation of Hsp90 function is also known as the Hsp90 “Chaperone Code.” Here, we demonstrate that only the active Mps1 is ubiquitinated on K86, K827, and K848 by the tumor suppressor von Hippel-Lindau (VHL) containing E3 enzyme, in a prolyl hydroxylation-independent manner and degraded in the proteasome. Furthermore, we show that this process regulates cell exit from the mitotic checkpoint. Collectively, our data demonstrates an interplay between the Hsp90 chaperone and VHL degradation machinery in regulating mitosis.     

Tumor suppressor Tsc1 is a new Hsp90 co‐chaperone that facilitates folding of kinase and non‐kinase clients

The tumor suppressors Tsc1 and Tsc2 form the tuberous sclerosis complex (TSC), a regulator of mTOR activity. Tsc1 stabilizes Tsc2; however, the precise mechanism involved remains elusive. The molecular chaperone heat‐shock protein 90 (Hsp90) is an essential component of the cellular homeostatic machinery in eukaryotes. Here, we show that Tsc1 is a new co‐chaperone for Hsp90 that inhibits its ATPase activity. The C‐terminal domain of Tsc1 (998–1,164 aa) forms a homodimer and binds to both protomers of the Hsp90 middle domain. This ensures inhibition of both subunits of the Hsp90 dimer and prevents the activating co‐chaperone Aha1 from binding the middle domain of Hsp90. Conversely, phosphorylation of Aha1‐Y223 increases its affinity for Hsp90 and displaces Tsc1, thereby providing a mechanism for equilibrium between binding of these two co‐chaperones to Hsp90. Our findings establish an active role for Tsc1 as a facilitator of Hsp90‐mediated folding of kinase and non‐kinase clients—including Tsc2—thereby preventing their ubiquitination and proteasomal degradation.     

The duration of the inhibitory effects with static stretching on quadriceps peak torque production

Although several studies have investigated the acute effect of static stretching exercises, the duration of exercises that negatively affects performance has not been ascertained. This study was conducted to determine the acute effect of different static stretching durations on quadriceps isometric and isokinetic peak torque production. The 50 participants were randomly allocated into five equivalent sized groups and were asked to perform a stretching exercise of different duration (no stretch, 10-second stretch, 20-second stretch, 30-second stretch, and 60-second stretch). The knee flexion range of motion and the isometric and concentric isokinetic peak torques of the quadriceps were measured before and after a static stretching exercise in the four experimental groups. The same parameters were examined in the control group (no stretch) without stretching, before and after a 5-minute passive rest. There were no significant differences among groups before the experimentation regarding their physical characteristics and performances (P > 0.05). These results reflect the different groups' homogeneity. Significant knee joint flexibility increases (P < 0.001) and significant isometric and isokinetic peak torque reductions (P < 0.05-0.001) have been shown to occur only after 30 and 60 seconds of quadriceps static stretching. Stretching reduced isometric peak torque by 8.5% and 16.0%, respectively. Concerning isokinetic peak torque after 30 and 60 seconds of stretching, it was reduced by 5.5% vs. 11.6% at 60 degrees/s and by 5.8% vs. 10.0% at 180 degrees/s. We suggest that torque decrements are related to changes of muscle neuromechanical properties. It is recommended that static stretching exercises of a muscle group for more than 30 seconds of duration be avoided before performances requiring maximal strength.     

Low avidity antibody: a reliable method to diagnose a recent HIV-1 infection

Standard serological tests (both EIA and Immunoblotting) have reached high levels of sensitivity and reproducibility, but do not indicate whether infection is recent or longstanding. Since many patients with HIV-1 infection are not usually diagnosed until symptom presentation, the possibility to distinguish between acute and chronic infection has become increasingly important for the purposes of therapeutic decision-making, partner notification and epidemiological surveillance. We evaluated a guanidine-based-antibody-avidity assay in a selected group of recent (within six months from seroconversion) and chronic (more than forty eight months) HIV-1 infections in an attempt to shed more light on the significance of the avidity index in establishing the time of infection. Sera from newly infected individuals showed a low mean avidity index (ranging from 0.35 to 0.60 with a standard deviation 0.09) at baseline and a clear increasing value at the following times of observation. Our data showed that an avidity index <0.70 might be presumptive of infection occurring within 9 months. Avidity index levels might distinguish between acute and chronic infection. The method is semi-automated, inexpensive and easy to perform, and estimates the time elapsed from seroconversion, thereby identifying a recent infection.     

Endoparasitoid wasp bracovirus-mediated inhibition of hemolin function and lepidopteran host immunosuppression

Successful embryonic development of parasitoid wasps in lepidopteran hosts is achieved through co-injection of polydna viruses whose gene products are thought to target the immune responses of the host. One gene product of the endosymbiont bracovirus of the parasitic wasp Cotesia rubecula, CrV1, has been reported to inhibit the immune responses of its endoparasitized lepidopteran host through interference with the haematocyte cytoskeletal structure. Here we establish that CcV1, the Cotesia congregata bracovirus orthologue of CrV1, is also uptaken by lepidopteran haemocytes and haemocyte-like established cell lines, but we also report on a different function of CcV1, which is highly relevant to the inhibition of the host immune responses and is based on its direct interaction with the pattern recognition molecule hemolin. Recombinant CcV1 inhibits hemolin functions, such as lipopolysaccharide binding and bacterial agglutination as well as bacterial phagocytosis by haemocytes and haemocyte-like cell lines, producing functional phenotypes equivalent to those observed to arise from RNAi-based inhibition of hemolin gene expression. Finally, we show that CcV1 and hemolin colocalize on the membrane surface of hemolin-expressing cells, a finding suggesting that CcV1 may be uptaken by haemocytes and inhibit haemocyte function as a result of its interaction with membrane-anchored hemolin.     

Effect of amino substitution on the excited state dynamics of uracil.

The excited state deactivation of two amino-substituted uracils, 5-aminouracil (5AU) and 6-aminouracil (6AU) in aqueous solution was studied by femtosecond fluorescence upconversion. The fluorescence of 6AU decays as fast as that of uracil with a unique time constant of about 100 femtoseconds. The fluorescence of 5AU exhibits a more complex behavior, fundamentally different from what we found in any other uracils: the decays are globally slower (up to several picoseconds) and depend strongly on the wavelength. This difference is attributed to the particular character of the amino group, affecting the out-of-plane motion of the 5-substituent which has been shown to be crucial for the ultrafast internal conversion occurring in uracils. Our observations indicate instead the formation of a transient fluorescent state which in turn is deactivated by a different relaxation process specific to the amino group.     

Synthesis and biological evaluation of novel angular fused Pyrrolocoumarins

Angular pyrrolocoumarins were synthesized from the reaction of 4-hydroxyindole or 5-hydroxyindole with DMAD and PPh(3) and were tested for anti-inflammatory and antioxidant activity. These compounds significantly inhibited the carrageenin-induced paw edema (60.5%-73.4%) and have important scavenging activity. Although their interaction with the free stable radical DPPH is not high, compound 9 is the most potent (73.4%) in the in vivo experiment. Compound 7 seems to be a potent LOX inhibitor. An attempt was made to correlate the biological results with their structural characteristics and physicochemical parameters.     

Mapping abundance distribution of small pelagic species applying hydroacoustics and Co-Kriging techniques

Hydroacoustic technology provides ground tools for the estimation of abundance and spatial distribution of pelagic species. The final products of such surveys, the interpolated choropleth maps, are based on a Geostatistical analysis of the acoustic measurements to minimise, as much as possible, the interpolation error, and to quantify uncertainty. The current study is based on fisheries acoustic measurements and satellite images covering the sea area of Thermaikos Gulf over the years 1996, 1997 and 1998. Spatial interpolations describing the abundance and distribution of small pelagic species in the research area, as well as sea surface temperature (SST), Chlorophyll-a content (SSC) and average depth, were produced, based on Ordinary and Universal Kriging and Co-Kriging Geostatistical methods. The results of the Geostatistical analysis showed that the Co-Kriging spatial interpolation method produced the best results regarding fish abundance when SST and average depth variables were included in the model. The latter indicates that there is an existing spatial cross-correlation between fish abundance and the environmental variables. Consequently, the potential reduction of the overall error in the estimation process, as presented in this study, is very significant, particularly with regard to error reduction in stock assessment and management. Guest editor: V. D. Valavanis Essential Fish Habitat Mapping in the Mediterranean