Translational regulation of PGHS-1 mRNA: 5' untranslated region and first two exons conferring negative regulation.

Prostaglandin endoperoxide H synthase-1 gene expression is described as inducible in a few contexts such as differentiation of megakaryoblastic MEG-01 cells into platelet-like structures. In the MEG-01 cells model of PGHS-1 gene induction, we previously reported a delay in protein synthesis and identified the translational step of gene expression as being regulated. In the current study, we mapped PGHS-1 mRNA sequences regulating translational efficiency and identified an RNA binding protein. The 5'UTR and first two exons of the PGHS-1 5' mRNA decreased the synthesis of Luciferase protein by approximately 80% without significant changes in mRNA levels when compared to controls. Both the PGHS-1 5'-UTR and the first two exons were required for activity. Sucrose density gradient fractionations of cytoplasmic extracts from MEG-01 cells infected with reporter constructs, either controls or containing PGHS-1 sequence, presented a similar profile of distribution of reporter transcripts between polysomal and non-polysomal fractions. RNA/protein interaction studies revealed nucleolin binding to the 135 nt PGHS-1 sequence. Mutation of the two NRE elements located in the 5'end of PGHS-1 mRNA sequence partially reduced the negative activity of the 135 nt sequence. Stable secondary structures predicted at the 5' end of the transcript are potentially involved in translational regulation. We propose that the 5'end of PGHS-1 mRNA represses translation and could delay the synthesis of PGHS-1 enzyme.

Artemis phosphorylated by DNA-dependent protein kinase associates preferentially with discrete regions of chromatin.

Artemis is a nuclear phosphoprotein required for genomic integrity whose phosphorylation is increased subsequent to DNA damage. Artemis phosphorylation by the DNA-dependent protein kinase (DNA-PK) and the association of Artemis with DNA-PK catalytic subunit (DNA-PKcs) have been proposed to be crucial for the variable, diversity, joining (V(D)J) reaction, genomic stability and cell survival in response to double-stranded DNA breaks. The exact nature of the effectors of Artemis phosphorylation is presently being debated. Here, we have delimited the interface on Artemis required for its association with DNA-PKcs and present the characterization of six DNA-PK phosphorylation sites on Artemis whose phosphorylation shows dependence on its association with DNA-PKcs and is induced by double-stranded DNA damage. Surprisingly, DNA-PKcs Artemis association appeared to be dispensable in a V(D)J recombination assay with stably integrated DNA substrates. Phosphorylation at two of the sites on Artemis, S516 and S645, was verified in vivo using phosphospecific antibodies. Basal Artemis S516 and S645 phosphorylation in vivo showed a significant dependence on DNA-PKcs association. However, regardless of its association with DNA-PKcs, phosphorylation of Artemis at both S516 and S645 was stimulated in response to the double-stranded DNA-damaging agent bleomycin, albeit to a lesser extent. This suggests that additional factors contribute to promote DNA damage-induced Artemis phosphorylation. Intriguingly, pS516/pS645 Artemis was concentrated in chromatin-associated nuclear foci in naïve cells. These foci were maintained upon DNA damage but failed to overlap with the damage-induced gammaH2AX. These results provide the expectation of a specific role for DNA-PK-phosphorylated Artemis in both naïve and damaged cells.

Novel Measures to Assess the Effects of Partial Sleep Deprivation on Sensory, Working, and Permanent Memory.

Sleepiness has repeatedly been demonstrated to affect performance on a variety of cognitive tasks. While the effects of total sleep deprivation (TSD) have been extensively studied, acute partial sleep deprivation (PSD), a more frequent form of sleep loss, has been studied much less often. The present study examined the effects of sleep deprivation on novel tasks involving classic sensory, working, and permanent memory systems. While the tasks did implicate different memory systems, they shared a need for effortful, sustained attention to maintain successful performance. Because of the novelty of the tasks, an initial study of the effects of TSD was carried out. The effects of PSD were subsequently examined in a second study, in which subjects were permitted only 4 h of sleep. A general detrimental effect of both total and PSD on accuracy of detection was observed and to a lesser extent, a slowing of the speed of responding on the different tasks. This overall effect is best explained by the often-reported inability to sustain attention following sleep loss. Specific effects on distinct cognitive processes were also observed, and these were more apparent following total than PSD.

Comparative optimism among drivers: an intergenerational portrait.

We describe a large cross-sectional study examining comparative optimism (CO) regarding the risk of car crash in three age cohorts (17-26, 27-64, 65 and older). The participants completed a questionnaire in which they were asked to indicate their personal risk of being in an at-fault crash over nine driving related events in comparison to young drivers, mid-aged drivers, and older drivers. The results indicated that all three age groups exhibited CO, but that the level varied according to the age group on which they had to base their comparative judgment. Interestingly, greater CO was constantly observed when the three age groups compared themselves to older drivers. Also, young males exhibited greater CO than female of the same age group.