Bioassaying Putative RNA-Binding Motifs in a Protein Encoded by a Gene That Influences Courtship and Visually Mediated Behavior in Drosophila: In Vitro Mutagenesis of Nona

The no-on-transient-A (nonA) gene of Drosophila melanogaster influences vision, courtship song, and viability. The nonA-encoded polypeptide is inferred to bind single-stranded nucleic acids. Although sequence-analysis of NONA implies that it belongs to a special interspecific family of this protein type, it does contain two classical RNA recognition motifs (RRM). Their behavioral significance was assayed by generating transgenic strains that were singly or multiply mutated within the relatively N-terminal motif (RRM1) or within RRM2. Neither class of mutation affected NONA binding to polytene chromosomes. The former mutations led to extremely low viability, accompanied by diminished adult longevities that were much worse than for a nonA-null mutant, implying that faulty interpolypeptide interactions might accompany the effects of the amino-acid substitutions within RRM1. All in vitro-mutated types caused optomotor blindness and an absence of transient spikes in the electroretinogram. Courtship analysis discriminated between the effects of the mutations: the RRM2-mutated type generated song pulses and trains that tended to be mildly mutant. These phenotypic abnormalities reinforce the notion that nonA''s ubiquitous expression has its most important consequences in the optic lobes, the thoracic ganglia, or both, depending in part on the nonA allele.     

Lytic phages obscure the cost of antibiotic resistance in Escherichia coli

The long-term persistence of antibiotic-resistant bacteria depends on their fitness relative to other genotypes in the absence of drugs. Outside the laboratory, viruses that parasitize bacteria (phages) are ubiquitous, but costs of antibiotic resistance are typically studied in phage-free experimental conditions. We used a mathematical model and experiments with Escherichia coli to show that lytic phages strongly affect the incidence of antibiotic resistance in drug-free conditions. Under phage parasitism, the likelihood that antibiotic-resistant genetic backgrounds spread depends on their initial frequency, mutation rate and intrinsic growth rate relative to drug-susceptible genotypes, because these parameters determine relative rates of phage-resistance evolution on different genetic backgrounds. Moreover, the average cost of antibiotic resistance in terms of intrinsic growth in the antibiotic-free experimental environment was small relative to the benefits of an increased mutation rate in the presence of phages. This is consistent with our theoretical work indicating that, under phage selection, typical costs of antibiotic resistance can be outweighed by realistic increases in mutability if drug resistance and hypermutability are genetically linked, as is frequently observed in clinical isolates. This suggests the long-term distribution of antibiotic resistance depends on the relative rates at which different lineages adapt to other types of selection, which in the case of phage parasitism is probably extremely common, as well as costs of resistance inferred by classical in vitro methods.     

Potentiation of synaptic responses in slices from the chick forebrain.

Coronal slices, containing part of the medial hyperstriatum ventrale (MHV), were cut from the left forebrains of domestic chicks and maintained in vitro. Records were made of the field responses evoked in the MHV by local electrical stimuli provided at 0.1 Hz. Two 1 min periods of stimulation at 5 Hz, separated by 10 min, were used in attempts to induce a persistent increase in the size of the postsynaptic response to test stimulation at 0.1 Hz. This procedure produced a potentiation which usually lasted longer than 2 h. The probability of inducing this persistent potentiation of the response (PPR) is not distributed evenly over the whole anteroposterior length of the MHV but is higher in slices that also contain the septo-mesencephalic tract ventrally. These are the slices that contain the intermediate part of the medial hyperstriatum ventrale (IMHV); an area that is essential for early behavioural learning. At this level PPR is not confined to the IMHV. It can also be produced in the lateral neostriatum in response to similar local stimulation at 5 Hz. No PPR was observed in either the caudal ectostriatum, or the paleostriatum.     

FBXO44 promotes DNA replication-coupled repetitive element silencing in cancer cells

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