The p53 isoforms are differentially modified by Mdm2

The discovery that the single p53 gene encodes several different p53 protein isoforms has initiated a flurry of research into the function and regulation of these novel p53 proteins. Full-length p53 protein level is primarily regulated by the E3-ligase Mdm2, which promotes p53 ubiquitination and degradation. Here, we report that all of the novel p53 isoforms are ubiquitinated and degraded to varying degrees in an Mdm2-dependent and -independent manner, and that high-risk human papillomavirus can degrade some but not all of the novel isoforms, demonstrating that full-length p53 and the p53 isoforms are differentially regulated. In addition, we provide the first evidence that Mdm2 promotes the NEDDylation of p53β. Altogether, our data indicates that Mdm2 can distinguish between the p53 isoforms and modify them differently.     

The analysis of recombinant interleukin-2 by thermospray liquid chromatography-mass spectrometry

The complete sequence of recombinant human interleukin-2 expressed in Escherichia coli has been confirmed by thermospray liquid chromatography-mass spectrometry (TS-LC-MS) of a tryptic digest derived from 100 micrograms (7 nmol) of reduced carboxymethylated interleukin-2. The preparation was shown by this method to contain predominantly unprocessed N-terminal initiator Met, with some authentic N-terminal Ala; the rest of the protein was as predicted from the DNA sequence, though some deamidated material was noted. TS-LC-MS proved to be a rapid and efficient method for surveying the protein tryptic peptide products allowing all the data to be collected in one chromatographic run; all tryptic fragments were identified by their molecular ions including those for the larger peptides (Mr 1500-3500) which, due to the presence of doubly and triply charged molecular ions, were brought within the mass range of the instrument (1800 Da). It is proposed that TS-LC-MS is a good general method for analyzing recombinant protein digests with respect to sequence confirmation, processing, and post-translational modification, and since each chromatographic peak is identified allows for subsequent monitoring of the protein by LC using uv detection. The method suffers from the disadvantages that all the sample is consumed during the experiment and that no fragment (sequence) ions are generally observed.     

Behavioural modification of a social spider by a parasitoid wasp

1. Manipulation of host behaviour by parasitoids has long captured the imagination of ecologists. Parasitoid wasps in the Polysphincta group of genera develop as external parasitoids of spiders. 2. In the present study, the previously undescribed interaction between a Zatypota sp. wasp (Ichneumonidae) and a social spider Anelosimus eximius (Theridiidae) is described. The larva of this Zatypota wasp is found to induce its host to disperse from their communal web and build an entirely enclosed web consisting of densely spun silk. 3. The wasp is observed to target primarily immature A. eximius individuals, with 37.5–44% of nests in a given area being parasitised. Of those nests, approximately 1.3–2.0% of individuals are hosts to the parasitoid larvae. Larger spider colonies had a significantly higher probability of harbouring parasitoids. 4. This interaction results in unusual behaviours for A. eximius induced by the parasitoid: (i) leaving the protection of the social nest and (ii) building a unique, altered web that it would not otherwise build. It is suggested that the wasp may be tapping into ancestral dispersal behaviours in its host and that a social species provides this wasp an evolutionary advantage by allowing a stable host source.     

The effect of age on enolase turnover in the free-living nematode, Turbatrix aceti.

The rates of synthesis and degradation of enolase and total soluble proteins slow with age in the free-living nematode, Turbatrix aceti. The half-lives are 73 and 58 h for soluble protein and enolase, respectively, in young organisms (5 days old). The respective figures are 163 and 161 h for old organisms (22–30 days old). Similar slowing of protein turnover occurs when the organisms are aged by a repeated screening procedure which avoids the use of fluorodeoxyuridine, an inhibitor of DNA synthesis normally added to aging cultures to obtain synchrony. The results support the idea that slowed protein turnover may be responsible for the formation of altered enzymes in old organisms.     

A Bird’s Eye View of Discard Reforms: Bird-Borne Cameras Reveal Seabird/Fishery Interactions

Commercial capture fisheries produce huge quantities of offal, as well as undersized and unwanted catch in the form of discards. Declines in global catches and legislation to ban discarding will significantly reduce discards, but this subsidy supports a large scavenger community. Understanding the potential impact of declining discards for scavengers should feature in an eco-system based approach to fisheries management, but requires greater knowledge of scavenger/fishery interactions. Here we use bird-borne cameras, in tandem with GPS loggers, to provide a unique view of seabird/fishery interactions. 20,643 digital images (one min⁻¹) from ten bird-borne cameras deployed on central place northern gannets Morus bassanus revealed that all birds photographed fishing vessels. These were large (>15 m) boats, with no small-scale vessels. Virtually all vessels were trawlers, and gannets were almost always accompanied by other scavenging birds. All individuals exhibited an Area-Restricted Search (ARS) during foraging, but only 42% of ARS were associated with fishing vessels, indicating much ‘natural’ foraging. The proportion of ARS behaviours associated with fishing boats were higher for males (81%) than females (30%), although the reasons for this are currently unclear. Our study illustrates that fisheries form a very important component of the prey-landscape for foraging gannets and that a discard ban, such as that proposed under reforms of the EU Common Fisheries Policy, may have a significant impact on gannet behaviour, particularly males. However, a continued reliance on ‘natural’ foraging suggests the ability to switch away from scavenging, but only if there is sufficient food to meet their needs in the absence of a discard subsidy.     

Do pollen beetles need pollen? The effect of pollen on oviposition,survival, and development of a flower‐feeding herbivore

Abstract.  1. Pollen is considered to be an important dietary component for many species of flower-feeding herbivores. Its influence on oviposition site selection by the pollen beetle Meligethes aeneus , and on the development of its larvae was investigated.
2. The effects of pollen presence and absence on adult, egg, and larval incidence in the field, and on larval development in the laboratory were compared through the use of Synergy, a composite hybrid oilseed rape Brassica napus variety comprising male-fertile (with pollen) and male-sterile (without pollen) plants.
3. In the field, adult females were more abundant on male-fertile plants during flowering, and a greater proportion of male-fertile than male-sterile buds were accepted for oviposition. These data indicate a possible role of pollen in oviposition site selection by female pollen beetles.
4. The numbers of first instar larvae on the two plant lines did not differ; however, more second instars were found on male-fertile than on male-sterile flowers. This suggests a greater larval survival on male-fertile plants, possibly due to the more readily available food resources and better nutrition afforded by the presence of pollen.
5. Laboratory experiments confirmed that a diet which included pollen improved survival to adulthood and resulted in heavier pupae and adults; however, pollen was not obligatory for larval survival and development.
6. The pollen beetle, previously thought to be an obligate pollen feeder, is therefore more generalist in its requirements for development. These findings may relate to the nutritional and behavioural ecology of other flower-feeding herbivores.     

Contraction of filaments of Escherichia coli after disruption of cell membrane by detergent.

The osmotic pressure within a living bacterium creates stresses in the peptidoglycan that stretch the sacculus. We measured the amount of stretch by monitoring the shrinkage of growing cells of Escherichia coli after removal of the osmotic pressure by disruption of the phospholipid membranes with sodium dodecyl sulfate. Because the rods of the wild type are so short, length changes of filaments of longer than 7 microns were measured on phase-contrast micrographs. The filaments were prepared by growing ftsA and ftsI strains under permissive conditions in rich medium and then shifting them to 42 degrees C for 40 to 180 min. During this time, the mutant cells became elongated but did not divide. The growing filaments were mounted on a glass surface that had been treated with poly-L-lysine or RNase. The filaments were photographed before being treated with sodium dodecyl sulfate. The filaments were rephotographed at the time when the first change in phase contrast was noted. Some filaments were also measured at 10-min time intervals from 0 to 60 min. The reduction in phase contrast signaled the leakage of solutes and the loss of turgor pressure. The average length of the filaments decreased 17%. If the circumference were stretched to the same degree, then the surface area in vivo would be 45% greater than in the relaxed state. For comparison, a fully cross-linked monolayer of E. coli peptidoglycan in its most compact conformation could stretch up to 300% in achieving the most extended conformation possible without splitting covalent bonds.     

Campylobacter pylori, the spiral bacterium associated with human gastritis, is not a true Campylobacter sp.

Comparison of partial 16S rRNA sequences from representative Campylobacter species indicates that the Campylobacter species form a previously undescribed basic eubacterial group, which is related to the other major groups only by very deep branching. This analysis was extended to include the spiral bacterium associated with human gastritis, Campylobacter pylori (formerly Campylobacter pyloridis). The distance between C. pylori and the other Campylobacter species is sufficient to exclude the pyloric organism from the Campylobacter genus. The results indicate that C. pylori is more closely related to Wolinella succinogenes than it is to the other Campylobacter species inspected. Another close relative of the campylobacters was found to be Thiovulum, a sulfide-dependent marine bacterium.