Enzymatic catalysis of anti-Baldwin ring closure in polyether biosynthesis

Polycyclic polyether natural products have fascinated chemists and biologists alike owing to their useful biological activity, highly complex structure and intriguing biosynthetic mechanisms. Following the original proposal for the polyepoxide origin of lasalocid and isolasalocid and the experimental determination of the origins of the oxygen and carbon atoms of both lasalocid and monensin, a unified stereochemical model for the biosynthesis of polyether ionophore antibiotics was proposed. The model was based on a cascade of nucleophilic ring closures of postulated polyepoxide substrates generated by stereospecific oxidation of all-trans polyene polyketide intermediates. Shortly thereafter, a related model was proposed for the biogenesis of marine ladder toxins, involving a series of nominally disfavoured anti-Baldwin, endo-tet epoxide-ring-opening reactions. Recently, we identified Lsd19 from the Streptomyces lasaliensis gene cluster as the epoxide hydrolase responsible for the epoxide-opening cyclization of bisepoxyprelasalocid A to form lasalocid A. Here we report the X-ray crystal structure of Lsd19 in complex with its substrate and product analogue to provide the first atomic structure-to our knowledge-of a natural enzyme capable of catalysing the disfavoured epoxide-opening cyclic ether formation. On the basis of our structural and computational studies, we propose a general mechanism for the enzymatic catalysis of polyether natural product biosynthesis.     

A plant pathogen virulence factor inhibits the eukaryotic proteasome by a novel mechanism

Pathogenic bacteria often use effector molecules to increase virulence. In most cases, the mode of action of effectors remains unknown. Strains of Pseudomonas syringae pv. syringae (Pss) secrete syringolin A (SylA), a product of a mixed non-ribosomal peptide/polyketide synthetase, in planta. Here we identify SylA as a virulence factor because a SylA-negative mutant in Pss strain B728a obtained by gene disruption was markedly less virulent on its host, Phaseolus vulgaris (bean). We show that SylA irreversibly inhibits all three catalytic activities of eukaryotic proteasomes, thus adding proteasome inhibition to the repertoire of modes of action of virulence factors. The crystal structure of the yeast proteasome in complex with SylA revealed a novel mechanism of covalent binding to the catalytic subunits. Thus, SylA defines a new class of proteasome inhibitors that includes glidobactin A (GlbA), a structurally related compound from an unknown species of the order Burkholderiales, for which we demonstrate a similar proteasome inhibition mechanism. As proteasome inhibitors are a promising class of anti-tumour agents, the discovery of a novel family of inhibitory natural products, which we refer to as syrbactins, may also have implications for the development of anti-cancer drugs. Homologues of SylA and GlbA synthetase genes are found in some other pathogenic bacteria, including the human pathogen Burkholderia pseudomallei, the causative agent of melioidosis. It is thus possible that these bacteria are capable of producing proteasome inhibitors of the syrbactin class.     

Arthropod community dynamics in undisturbed and intensively managed mountain brush habitats

The population dynamics and decomposition activities of litter arthropods in an unmanaged sagehrush/bitterbrush habitat in southeastern Wyoming were assessed in 1986. The effects of sagebrush/bitterbrush management practices on litter arthropod communities and the role of these communities in decomposition were also assessed. Insecticide applications were used to selectively exclude arthropods in order to determine the ecological impact of these detritivores. Application of a herbicide, 2,4-D, was associated with increased arthropod populations for 30 days following treatment. Mowing increased arthropod densities at two different times, 10-30 days and 50 days post treatment, perhaps as a result of functional and numerical responses by litter arthropods. Elimination of arthropods from otherwise undisturbed shrub habitats by the use of broad-spectrum insecticides reduced the rate of litter decomposition during the growing season, indicating that these organisms play an important role in decomposition and nutrient cycling.     

Catastrophic wildfire and number of populations as factors influencing risk of extinction for Gila trout ( Oncorhynchus gilae )

We used the computer program RAMAS to explore the sensitivity of an extinction-risk model for the Gila trout ( Oncorhynchus gilae ) to management of wildfires and number of populations of the species. The Gila trout is an endangered salmonid presently restricted to very few headwaters of the Gila and San Francisco river tributaries in southwestern New Mexico. Life history data for 10 extant populations were used to examine sensitivity of the species viability to changes in a variety of factors including population size, fecundity, life stage structure, number of populations, severity and probability of forest fires, and a regulated fishery. The probability and severity of forest fires and number of populations had the greatest effect on viability. Results indicate that successful conservation of Gila trout requires establishment of additional populations and reduction of the severity of forest fires through a program incorporating more frequent, but less severe, fires.