Campylobacter jejuni fatty acid synthase II: Structural and functional analysis of β-hydroxyacyl-ACP dehydratase (FabZ)

Fatty acid biosynthesis is crucial for all living cells. In contrast to higher organisms, bacteria use a type II fatty acid synthase (FAS II) composed of a series of individual proteins, making FAS II enzymes excellent targets for antibiotics discovery. The β-hydroxyacyl-ACP dehydratase (FabZ) catalyzes an essential step in the FAS II pathway. Here, we report the structure of Campylobacter jejuni FabZ (CjFabZ), showing a hexamer both in crystals and solution, with each protomer adopting the characteristic hot dog fold. Together with biochemical analysis of CjFabZ, we define the first functional FAS II enzyme from this pathogen, and provide a framework for investigation on roles of FAS II in C. jejuni virulence.     

Molecular phylogeny and host-plant use (Lamiaceae) of the Thymogethes pollen beetles (Coleoptera)

The 24 members of the Euro-Asiatic genus Thymogethes are highly specialized pollen beetles associated as larvae with flowers of Lamiaceae Nepetoideae. All members of the genus were analysed in within the framework of an integrative taxonomy approach, which was aimed to reconstruct the phylogenetic relationships, as well as the possible pattern of evolution of their larval-host-plant association. Evidence from multiple molecular markers [COI; 16S; H3], combined with an estimation of divergence times using an average rate of 0.0177 substitutions/site/My among branches, placed the origin of the genus at a minimum of 9–10 Mya. This date of origin approximates the known evolution of the host plants in Euro-Mediterranean areas. Evidence from combined molecular and cladistic morphological analyses resulted in suitable agreement with the previously established morphology-based systematics of the genus, although members of the exilis species-group were split into three clades. The only disagreement between results of this new combined phylogeny and previous classification is in the exclusion of “Thymogethes” grenieri. This species is herein positioned outside the genus, based on molecular evidence. Our analysis depicts several Thymogethes species differentiating in the last few Mys, specifically those included in the T. lugubris species-group. Combined evidence from DNA, morphology and ancestral state parsimony reconstruction of larval-host-plant associations suggests that subtribe Menthinae likely represents the ancestral host plants, with a series of independent host shifts during the radiation of the clade, in association first with Menthinae and subsequently with Lavandulinae and Nepetinae. Steno-oligophagy is the most frequent (86%) condition, while strictly monophagous species are less numerous (14%).     

Unrecognized diversity of a scale worm,Polyeunoa laevis (Annelida: Polynoidae), that feeds on soft coral

A goal of taxonomy is to employ a method of classification based on phylogeny that captures the morphological and genetic diversity of organismal lineages. However, morphological and genetic diversity may not always be concordant, leading to challenges in systematics. The scale worm Polyeunoa laevis has been hypothesized to represent a species complex based on morphology, although there is little knowledge of its genetic diversity. Commonly found in Antarctic waters and usually associated with gorgonian corals (especially Thouarella), this taxon is also reported from the south-west Atlantic, Magellanic and sub-Antarctic regions. We employ an integrative taxonomic approach to examine the traditional morphological characters used for scale worm identification in combination with COI mitochondrial gene data and whole mtDNA genomes. Moreover, we consider P. laevis's association with Thouarella by examining data from the mMutS gene, a soft-coral phylogenetic marker. Analyses for P. laevis recovered 3 clades, two in Antarctic waters and one from the Argentina-Indian Ocean. Interestingly, genetic and morphological results show differences between specimens from South Argentina and the Antarctic region, suggesting that open ocean barriers might have limited gene flow from these regions. Bayesian phylogenetic analyses for Thouarella resulted in at least 12 lineages, although some of the lineages consist of only a single individual. Our results show different evolutionary histories for both species, confirming that association between these scale worms and their hosts is not restrictive. For both taxonomic groups, biodiversity in the Southern Ocean appears to be underestimated.     

Potent inhibitors of HCV-NS3 protease derived from boronic acids

Chronic hepatitis C infection is the leading causes for cirrhosis of the liver and hepatocellular carcinoma, leading to liver failure and liver transplantation. The etiological agent, HCV virus produces a single positive strand of RNA that is processed with the help of serine protease NS3 to produce mature virus. Inhibition of NS3 protease can be potentially used to develop effective drugs for HCV infections. Numerous efforts are now underway to develop potent inhibitors of HCV protease that contain ketoamides as serine traps. Herein we report the synthesis of a series of potent inhibitors that contain a boronic acid as a serine trap. The activity of these compounds were optimized to 200 pM. X-ray structure of compound 17 bound to NS3 protease is also discussed.