Identification of determinants involved in initiation of hepatitis C virus RNA synthesis by using intergenotypic replicase chimeras

The 5' nontranslated region (NTR) and the X tail in the 3' NTR are the least variable parts of the hepatitis C virus (HCV) genome and play an important role in the initiation of RNA synthesis. By using subgenomic replicons of the HCV isolates Con1 (genotype 1) and JFH1 (genotype 2), we characterized the genotype specificities of the replication signals contained in the NTRs. The replacement of the JFH1 5' NTR and X tail with the corresponding Con1 sequence resulted in a significant decrease in replication efficiency. Exchange of the X tail specifically reduced negative-strand synthesis, whereas substitution of the 5' NTR impaired the generation of progeny positive strands. In search for the proteins involved in the recognition of genotype-specific initiation signals, we analyzed recombinant nonstructural protein 5B (NS5B) RNA polymerases of both isolates and found some genotype-specific template preference for the 3' end of positive-strand RNA in vitro. To further address genotype specificity, we constructed a series of intergenotypic replicon chimeras. When combining NS3 to NS5A of Con1 with NS5B of JFH1, we observed more-efficient replication with the genotype 2a X tail, indicating that NS5B recognizes genotype-specific signals in this region. In contrast, a combination of the NS3 helicase with NS5A and NS5B was required to confer genotype specificity to the 5' NTR. These results present the first genetic evidence for an interaction between helicase, NS5A, and NS5B required for the initiation of RNA synthesis and provide a system for the specific analysis of HCV positive- and negative-strand syntheses.     

Sea turtles: navigating with magnetism

Young sea turtles use the Earth's magnetic field as a source of navigational information during their epic transoceanic migrations and while homing. A new study using satellite telemetry has now demonstrated for the first time that adult turtles also navigate using the Earth's magnetic field.     

Biogeography and evolution of Dolichandra (Bignonieae,Bignoniaceae)

Little information on evolutionary relationships of Neotropical organisms or on the factors that have shaped the diversity currently encountered in this region is available. However, it is clear that biotic interactions and abiotic aspects have played important roles for species diversification in the region. This study focuses on Dolichandra (Bignonieae, Bignoniaceae), a clade of Neotropical lianas that is distributed broadly across different habitats and with diverse pollination and dispersal systems. We used sequences from two plastid DNA markers (ndhF and rpl32‐trnL) and one nuclear gene (PepC) to infer phylogenetic relationships in Dolichandra using parsimony and Bayesian approaches. We then used this phylogenetic framework as basis to study the biogeographic history, reconstruct the evolution of morphological characters and test the impact of morphology and environment on the diversification of the genus. More specifically, we: (1) time‐calibrate the phylogenetic tree of Dolichandra; (2) estimate the ancestral areas of the various lineages; (3) estimate the ancestral states of discrete and continuous morphological traits; (4) test for phylogenetic signal in environmental and phenotypic data; and (5) test whether morphological characters and/or niche evolution are correlated with cladogenesis. All Dolichandra spp. are monophyletic in the combined molecular phylogeny; relationships among species are generally well resolved, although poorly supported in some instances. The genus is inferred to have originated 36.43–26.23 Mya, possibly in eastern South America. Ancestral state reconstructions of continuous and discrete floral characters inferred a mixed morphology as the ancestral condition for the group. Phylogenetic signal differed between perianth and sexual whorls and gradual evolution was recovered for all traits except style length and anther length. Environmental variables showed no phylogenetic signal and a pattern of variation that was not correlated with branch length, suggesting that environmental transitions were concomitant with speciation. Dispersal is inferred to be the main driver of the differential distribution observed among species. In addition, climatic preferences and floral characters seem to have been important reproductive barriers in Dolichandra. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 403–420.     

Activity-dependent clustering of functional synaptic inputs on developing hippocampal dendrites

During brain development, before sensory systems become functional, neuronal networks spontaneously generate repetitive bursts of neuronal activity, which are typically synchronized across many neurons. Such activity patterns have been described on the level of networks and cells, but the fine-structure of inputs received by an individual neuron during spontaneous network activity has not been studied. Here, we used calcium imaging to record activity at many synapses of hippocampal pyramidal neurons simultaneously to establish the activity patterns in the majority of synapses of an entire cell. Analysis of the spatiotemporal patterns of synaptic activity revealed a fine-scale connectivity rule: neighboring synapses (<16?μm intersynapse distance) are more likely to be coactive than synapses that are farther away from each other. Blocking spiking activity or NMDA receptor activation revealed that the clustering of synaptic inputs required neuronal activity, demonstrating a role of developmentally expressed spontaneous activity for connecting neurons with subcellular precision.