Review: postchaperonin tubulin folding cofactors and their role in microtubule dynamics

The microtubule cytoskeleton consists of a highly organized network of microtubule polymers bound to their accessory proteins: microtubule-associated proteins, molecular motors, and microtubule-organizing proteins. The microtubule subunits are heterodimers composed of one alpha-tubulin polypeptide and one beta-tubulin polypeptide that should undergo a complex folding processing before they achieve a quaternary structure that will allow their incorporation into the polymer. Due to the extremely high protein concentration that exists at the cell cytoplasm, there are alpha- and beta-tubulin interacting proteins that prevent the unwanted interaction of these polypeptides with the surrounding protein pool during folding, thus allowing microtubule dynamics. Several years ago, the development of a nondenaturing electrophoretic technique made it possible to identify different tubulin intermediate complexes during tubulin biogenesis in vitro. By these means, the cytosolic chaperonin containing TCP-1 (CCT or TriC) and prefoldin have been demonstrated to intervene through tubulin and actin folding. Various other cofactors also identified along the alpha- and beta-tubulin postchaperonin folding route are now known to have additional roles in tubulin biogenesis such as participating in the synthesis, transport, and storage of alpha- and beta-tubulin. The future characterization of the tubulin-binding sites to these proteins, and perhaps other still unknown proteins, will help in the development of chemicals that could interfere with tubulin folding and thus modulating microtubule dynamics. In this paper, current knowledge of the above postchaperonin folding cofactors, which are in fact chaperones involved in tubulin heterodimer quaternary structure achievement, will be reviewed.     

Foraging movements of Eurasian griffon vultures (Gyps fulvus): implications for supplementary feeding management

The outbreak of bovine spongiform encephalopathy provoked restrictive European sanitary legislation that forced farmers to remove livestock carcasses from the wild. This had serious repercussions for the scavenger raptor guild. Against this background, we developed a study to analyse the foraging movements of Eurasian griffon vultures (Gyps fulvus) in northern Spain. We ringed 241 griffon vultures with alphanumeric plastic rings in Biscay between 2000 and 2011 and set experimental feeding stations in 24 sites over an area of 10,614 km²; recording re-sightings of the ringed vultures between 2005 and 2012. Using these re-sighting records, we tested whether birds randomly moved long distances whilst searching for food, or if vulture re-sightings were restricted to a few feeding sites within a limited area. We summarised 329 field-work days, with an average of 2.06 ringed vultures re-sighted per day, accounting for 1,017 re-sightings. Adult vultures were detected in three separate foraging nuclei within the study area. Movements out of the main foraging nuclei were statistically less frequent than would be expected if adult vultures accessed all resources at a similar rate. Once established at breeding areas, subadult vultures behaved in the same way as adults. Our results suggest that vultures’ home ranges are largely restricted to zones close to breeding areas. This has important consequences from a conservation point of view, suggesting that management decisions should take into consideration spatial scale effects.     

Ungulate Vehicle Collisions in a Peri-Urban Environment: Consequences of Transportation Infrastructures Planned Assuming the Absence of Ungulates

Ungulate vehicle collisions (UVC) provoke serious damage, including human casualties, and a large number of measures have been developed around the world to avoid collisions. We analyse the main factors involved in UVC in a road network built in the absence of ungulates, where mitigation structures to avoid UVC were not adequately considered. Ungulate population greatly increased during the last two decades and now Roe Deer and Wild Boars are widely distributed over the study area, but even after this increase, the road network was not adapted to avoid UVC. A total of 235 Roe Deer (RDVC) and 153 Wild Boar vehicle collisions (WBVC) were recorded between January 2008 and December 2011. We randomly selected 289 sample points (87 RDVC, 60 WBVC and 142 controls) separated by at least 500 metres from the next closest point and measured 19 variables that could potentially influence the vehicle collisions. We detected variations in the frequency of RDVC on a monthly basis, and WBVC was higher at weekends but no significant differences were detected on a monthly basis. UVC were more likely to occur at locations where sinuosity of the road, velocity, surface of shrub and deciduous forest area were greater, the presence of fences entered with positive relationship and distance to the nearest building was less. RDVC were more likely to occur at locations where timber forest area increased and distance to the nearest building decreased and WBVC was related to open fields cover and also to the presence of fences. Sinuosity and velocity entered in both cases as significant factors. Major roads, in which the traffic volume is greater and faster, caused more accidents with ungulates than secondary roads. Nowadays, the high frequency of ungulate road-kills deserves a new strategy in order to adapt infrastructure and adopt mitigation measures.     

A simple strategy for detecting moving objects during locomotion revealed by animal-robot interactions

An important role of visual systems is to detect nearby predators, prey, and potential mates, which may be distinguished in part by their motion. When an animal is at rest, an object moving in any direction may easily be detected by motion-sensitive visual circuits. During locomotion, however, this strategy is compromised because the observer must detect a moving object within the pattern of optic flow created by its own motion through the stationary background. However, objects that move creating back-to-front (regressive) motion may be unambiguously distinguished from stationary objects because forward locomotion creates only front-to-back (progressive) optic flow. Thus, moving animals should exhibit an enhanced sensitivity to regressively moving objects. We explicitly tested this hypothesis by constructing a simple fly-sized robot that was programmed to interact with a real fly. Our measurements indicate that whereas walking female flies freeze in response to a regressively moving object, they ignore a progressively moving one. Regressive motion salience also explains observations of behaviors exhibited by pairs of walking flies. Because the assumptions underlying the regressive motion salience hypothesis are general, we suspect that the behavior we have observed in Drosophila may be widespread among eyed, motile organisms.     

The IntI1 Integron Integrase Preferentially Binds Single-Stranded DNA of the attC Site

IntI1 integrase is a member of the prokaryotic DNA integrase superfamily. It is responsible for mobility of antibiotic resistance cassettes found in integrons. IntI1 protein, as well as IntI1-COOH, a truncated form containing its carboxy-terminal domain, has been purified. Electrophoretic mobility shift assays were carried out to study the ability of IntI1 to bind the integrase primary target sites attI and aadA1 attC. When using double-stranded DNA as a substrate, we observed IntI1 binding to attI but not to attC. IntI1-COOH did not bind either attI or attC, indicating that the N-terminal domain of IntI1 was required for binding to double-stranded attI. On the other hand, when we used single-stranded (ss) DNA substrates, IntI1 bound strongly and specifically to ss attC DNA. Binding was strand specific, since only the bottom DNA strand was bound. Protein IntI1-COOH bound ss attC as well as did the complete integrase, indicating that the ability of the protein to bind ss aadA1 attC was contained in the region between amino acids 109 and 337 of IntI1. Binding to ss attI DNA by the integrase, but not by IntI1-COOH, was also observed and was specific for the attI bottom strand, indicating similar capabilities of IntI1 for binding attI DNA in either double-stranded or ss conformation. Footprinting analysis showed that IntI1 protected at least 40 bases of aadA1 attC against DNase I attack. The protected sequence contained two of the four previously proposed IntI1 DNA binding sites, including the crossover site. Preferential ssDNA binding can be a significant activity of IntI1 integrase, which suggests the utilization of extruded cruciforms in the reaction mechanisms leading to cassette excision and integration.     

Regulation of lipid metabolism by dipyridamole and adenosine antagonists in rat adipocytes

1. Acute effects of dipyridamole, an inhibitor of adenosine transport, direct activators of adenylate cyclase and thirteen adenosine antagonist analogs on fatty acid synthesis have been examined in terms of the control of [1-14C]acetate incorporation into labeled fatty acids in the presence of glucose. 2. This monosaccharide acts as a stimulator of lipogenesis by generating NADPH for the lipid synthesis. 3. The relationship between lipogenesis and lipolysis was compared with a variety of adenylate cyclase stimulators. 4. The data obtained reveals that dipyridamole potentiated the inhibitory or stimulatory effects of isoproterenol and forskolin on lipogenesis and on lipolysis, respectively. 5. In these cases the data show that it exists an inverse relationship between lipogenesis and lipolysis. 6. Dipyridamole and methylxanthine analogs only moderately affect the rate of lipolysis whereas its effects are more potent on lipogenesis and lend further support to the hypothesis that dipyridamole antagonize adenosine actions as well as methyl xanthines. 7. These results suggest that dipyridamole and adenosine antagonists alter lipogenesis independently of the lipolytic process and that it exists an inverse relationship between lipogenesis and lipolysis under some conditions whereas there are not under others.     

Serotonin increases the cAMP concentration and the phosphoenolpyruvate carboxykinase mRNA in rat kidney, small intestine, and liver.

Within 60 min of the administration of serotonin to fasted-refed rats, there was a 5-, 16-, and 20-fold stimulation of the mRNA coding for the cytosolic form of P-enolpyruvate carboxykinase in the kidney, small intestine and liver, respectively. This stimulation was 5-, 1.3-, and 2-fold higher than noted in the same tissue after 24 h of starvation. Dose- and time-response curves to serotonin in the three tissues were similar. The level of PEPCK mRNA in the liver was significantly elevated within 30 min of serotonin administration, whereas 60 min was required in the small intestine and the kidney. The direct effect of serotonin on PEPCK mRNA was also assessed in hepatocytes maintained in primary culture. Serotonin (10(-8) M to 10(-4) M) caused a dose-dependent increase in the level of PEPCK mRNA and a transient increase in cAMP concentration. Within the first min of serotonin (10(-6) M) addition to cells, cAMP concentration increased 4-fold and returned after 10 min to basal level. Therefore, these results provide functional evidence of serotonin action in the rat peripheric tissues and suggest that cAMP is involved in its intracellular signalling.