Relative confribution of dehydrogenases to overall respiratory ETh activity in some marine organisms

Respiration is an oxidation-reduction process in which the electronflux through the respiratory electron transfer system (ETS)is sustained by the action of different dehydrogenases. Theseenzymes, as parts of the ETS, oxidize natural substrates (succinate,NADH and NADPH) of the cells and use the reducing equivalentsto activate ATP synthesis. We studied the relative contributionof the three main dehydrogenases to the overall ETh activityin some marine organisms. Each organism was analysed for thecombined and separate activities of NADH, NADPH and succinatedehydrogenases. The ETS activity was measured as the abilityof each organism to reduce the tetrazolium salt, INT, when suppliedwith their natural substrates. The results showed that (i) NADHdehydrogenase was generally the most active dehydrogenase inprokaryotic and eukaryotic cells; (ii) INT does not fully collectreducing equivalents from succinate through the succinate dehydrogenase;and (iii) the sum of the activities measured separately exceedsthe combined activity when the three enzymes are measured together.We suggest that competition of the individual dehydrogenasesfor a common limiting electron acceptor, ubiquinone, may explainthese observations.     

Development and evaluation of a core genome multilocus sequence typing scheme for Paenibacillus larvae,the deadly American foulbrood pathogen of honeybees

Paenibacillus larvae is the causative agent of the fatal American foulbrood disease in honeybees (Apis mellifera). Strain identification is vital for preventing the spread of the disease. To date, the most accessible and robust scheme to identify strains is the multilocus sequence typing (MLST) method. However, this approach has limited resolution, especially for epidemiological studies. As the cost of whole-genome sequencing has decreased and as it becomes increasingly available to most laboratories, an extended MLST based on the core genome (cgMLST) presents a valuable tool for high-resolution investigations. In this study, we present a standardized, robust cgMLST scheme for P. larvae typing using whole-genome sequencing. A total of 333 genomes were used to identify, validate and evaluate 2419 core genes. The cgMLST allowed fine-scale differentiation between samples that had the same profile using traditional MLST and allowed for the characterization of strains impossible by MLST. The scheme was successfully used to trace a localized Swedish outbreak, where a cluster of 38 isolates was linked to a country-wide beekeeping operation. cgMLST greatly enhances the power of a traditional typing scheme, while preserving the same stability and standardization for sharing results and methods across different laboratories.     

Rae1/YacP,a new endoribonuclease involved in ribosome-dependent mRNA decay in Bacillus subtilis

The PIN domain plays a central role in cellular RNA biology and is involved in processes as diverse as rRNA maturation, mRNA decay and telomerase function. Here, we solve the crystal structure of the Rae1 (YacP) protein of Bacillus subtilis, a founding member of the NYN (Nedd4-BP1/YacP nuclease) subfamily of PIN domain proteins, and identify potential substrates in vivo. Unexpectedly, degradation of a characterised target mRNA was completely dependent on both its translation and reading frame. We provide evidence that Rae1 associates with the B. subtilis ribosome and cleaves between specific codons of this mRNA in vivo. Critically, we also demonstrate translation-dependent Rae1 cleavage of this substrate in a purified translation assay in vitro. Multiple lines of evidence converge to suggest that Rae1 is an A-site endoribonuclease. We present a docking model of Rae1 bound to the B. subtilis ribosomal A-site that is consistent with this hypothesis and show that Rae1 cleaves optimally immediately upstream of a lysine codon (AAA or AAG) in vivo.     

New insigths on the metabolic diversity among the epibiotic microbial communitiy of the hydrothermal shrimp Rimicaris exoculata

The Rimicaris exoculata dominates the megafauna of some of the Mid Atlantic ridge hydrothermal vent sites. This species harbors a rich community of bacterial epibionts inside its gill chamber. Literature data indicate that a single 16S rRNA phylotype dominates this epibiotic community, and is assumed to be a sulfide-oxidizing bacteria. However attempts of cultivation were not successful and did not allow to confirm it. The aim of our study was to test the hypothesis of sulfide oxidation in the gill chamber, by a multidisciplinary approach, using in vivo experiments at in situ pressure in the presence of sulfide, microscopic observations and a molecular survey. Morphology of microorganisms, before and after treatment, was analyzed to test the effect of sulfide depletion and re-exposure. Our observations, as well as molecular data indicate a wider diversity than previously described for this shrimp's epibiotic community. We observed occurrence of bacterial intracellular sulfur- and iron-enriched granules and some methanotrophic-like bacteria cells for the first time. Genes that are characteristic of methane-oxidizing (pmoA) and sulfide-oxidizing (APS) bacteria were identified. These results suggest that three metabolic types (iron, sulfide and methane oxidation) may co-occur within the epibiont community associated with Rimicaris exoculata. As this shrimp colonizes chemically contrasted environments, the relative abundance of each metabolic type could vary according to the local availability of reduced compounds.     

Intracellular amorphous Ca-carbonate and magnetite biomineralization by a magnetotactic bacterium affiliated to the Alphaproteobacteria

Bacteria synthesize a wide range of intracellular submicrometer-sized inorganic precipitates of diverse chemical compositions and structures, called biominerals. Their occurrences, functions and ultrastructures are not yet fully described despite great advances in our knowledge of microbial diversity. Here, we report bacteria inhabiting the sediments and water column of the permanently stratified ferruginous Lake Pavin, that have the peculiarity to biomineralize both intracellular magnetic particles and calcium carbonate granules. Based on an ultrastructural characterization using transmission electron microscopy (TEM) and synchrotron-based scanning transmission X-ray microscopy (STXM), we showed that the calcium carbonate granules are amorphous and contained within membrane-delimited vesicles. Single-cell sorting, correlative fluorescent in situ hybridization (FISH), scanning electron microscopy (SEM) and molecular typing of populations inhabiting sediments affiliated these bacteria to a new genus of the Alphaproteobacteria. The partially assembled genome sequence of a representative isolate revealed an atypical structure of the magnetosome gene cluster while geochemical analyses indicate that calcium carbonate production is an active process that costs energy to the cell to maintain an environment suitable for their formation. This discovery further expands the diversity of organisms capable of intracellular Ca-carbonate biomineralization. If the role of such biomineralization is still unclear, cell behaviour suggests that it may participate to cell motility in aquatic habitats as magnetite biomineralization does.Subject terms: Phylogenetics, Biodiversity, Biogeochemistry, Water microbiology