Activation Domain-dependent Degradation of Somatic Wee1 Kinase

Cell cycle progression is dependent upon coordinate regulation of kinase and proteolytic pathways. Inhibitors of cell cycle transitions are degraded to allow progression into the subsequent cell cycle phase. For example, the tyrosine kinase and Cdk1 inhibitor Wee1 is degraded during G₂ and mitosis to allow mitotic progression. Previous studies suggested that the N terminus of Wee1 directs Wee1 destruction. Using a chemical mutagenesis strategy, we report that multiple regions of Wee1 control its destruction. Most notably, we find that the activation domain of the Wee1 kinase is also required for its degradation. Mutations in this domain inhibit Wee1 degradation in somatic cell extracts and in cells without affecting the overall Wee1 structure or kinase activity. More broadly, these findings suggest that kinase activation domains may be previously unappreciated sites of recognition by the ubiquitin proteasome pathway.     

Prevalence of Tree Regeneration by Sprouting and Seeding Along a Rainfall Gradient in Hawai'i

Drought stress in tropical dry forests is thought to result in greater asexual regeneration via vegetative sprouting ( e.g ., basal, root, and branch layering) than occurs in moister tropical forests. We tested this hypothesis by examining the prevalence of tree sprouting and seeding in tropical forests located along a rainfall gradient on the island of Hawai'i. Additionally, we examined the potential for novel disturbance, feral pig Sus scrofa rooting and trampling, to alter patterns in tree regeneration mode. We found greater sprouting (in terms of relative density and basal area) in dry forests than in mesic and wet forests, supporting the hypothesis. We also found that feral pig disturbance is negatively correlated with the relative density and basal area of seedlings in wet forests, but is positively correlated with the relative importance of sprouting, and the richness and diversity of sprouting species. Our results suggest rainfall regimes may be an important factor controlling broad-scale patterns in tree regeneration mode, and that exotic ungulates can significantly modify such patterns with potential consequences for the structure and dynamics of tree populations and communities.     

Structural analysis of Chi1 Chitinase from Yen-Tc: the multisubunit insecticidal ABC toxin complex of Yersinia entomophaga

Yersinia entomophaga MH96 is a native New Zealand soil bacterium that secretes a large ABC-type protein toxin complex, Yen-Tc, similar to those produced by nematode-associated bacteria such as Photorhabdus luminescens. Y. entomophaga displays an exceptionally virulent pathogenic phenotype in sensitive insect species, causing death within 72 h of infection. Because of this phenotype, there is intrinsic interest in the mechanism of action of Yen-Tc, and it also has the potential to function as a novel class of biopesticide. We have identified genes that encode chitinases as part of the toxin complex loci in Y. entomophaga MH96, P. luminescens, Photorhabdus asymbiotica and Xenorhabdus nematophila. Furthermore, we have shown that the secreted toxin complex from Y. entomophaga MH96 includes two chitinases as an integral part of the complex, a feature not described previously in other ABC toxins and possibly related to the severe disease caused by this bacterium. We present here the structure of the Y. entomophaga MH96 Chi1 chitinase, determined by X-ray crystallography to 1.74 Å resolution, and show that a ring of five symmetrically arranged lobes on the surface of the Yen-Tc toxin complex structure, as determined by single-particle electron microscopy, provides a good fit to the Chi1 monomer. We also confirm that the isolated chitinases display endochitinase activity, as does the complete toxin complex.     

Observations concerning the effects of radiations on the segregation of chromosomes

Nondisjunction of X and of fourth chromosomes was observed following the exposure of immature oocytes of Drosophila melanogaster to doses of X-radiation of from 1000 to 4000 R. No evidence for a threshold was found in this range for either kind of trisomy; this evidence alone does not exclude the possibility that one might be found at some lower dose. The mating of the treated females with males having an attached-XY chromosome permitted the recovery of fertile males that would otherwise have been XO and sterile. Testing of these showed some 22% to be triplo-4, having two maternal fourth chromosomes. Marking the left arm of chromosome 4 with a small duplication made it possible to score marker losses such as might result from interchange with another acrocentric (e.g., the X). There is a high coincidence of marker loss from chromosome 4 and both the XO and triplo-4 conditions, with the highest incidence of marker loss being when these have occurred together. The interpretation that the altered 4's are half-translocations resulting from X-4 interchange is further supported by the finding that they also show altered assortative behavior in compound-X females lacking a Y, when in combination with a standard fourth chromosome. A few show regular segregation from the attached-XY in the male, supporting the interpretation that they have the base of the X capped by the right arm of chromosome 4. It is argued that other trisomies may come about by mechanisms similar to that responsible for the triplo-4 condition. Furthermore, if rearrangement plays a part in the origin of trisomy, operating by altering division-I orientation as a result of heterologous conjunction maintained by chromatid interchange, it is unlikely that there will be a threshold for its induction.