Self-assembly of the general membrane-remodeling protein PVAP into sevenfold virus-associated pyramids

Viruses have developed a wide range of strategies to escape from the host cells in which they replicate. For egress some archaeal viruses use a pyramidal structure with sevenfold rotational symmetry. Virus-associated pyramids (VAPs) assemble in the host cell membrane from the virus-encoded protein PVAP and open at the end of the infection cycle. We characterize this unusual supramolecular assembly using a combination of genetic, biochemical, and electron microscopic techniques. By whole-cell electron cryotomography, we monitored morphological changes in virus-infected host cells. Subtomogram averaging reveals the VAP structure. By heterologous expression of PVAP in cells from all three domains of life, we demonstrate that the protein integrates indiscriminately into virtually any biological membrane, where it forms sevenfold pyramids. We identify the protein domains essential for VAP formation in PVAP truncation mutants by their ability to remodel the cell membrane. Self-assembly of PVAP into pyramids requires at least two different, in-plane and out-of-plane, protein interactions. Our findings allow us to propose a model describing how PVAP arranges to form sevenfold pyramids and suggest how this small, robust protein may be used as a general membrane-remodeling system.Release of virus particles from infected cells is the last essential step of the viral replication cycle. In the course of this process, virions face the challenging task of crossing the cell envelope. Viruses have developed an arsenal of diverse strategies to overcome this problem. Most bacterial viruses are lytic and induce lysis of the infected cell with help of the holin-endolysin system (1), whereas others disrupt the host cell envelope via inhibition of the murein biosynthesis pathway (2). The morphological and genomic properties of archaeal viruses (3) suggested that their egress from host cells may have unusual traits that are different from those of bacterial viruses. Indeed, although most archaeal viruses exit cells without lysis, some, in particular the Sulfolobus islandicus rod-shaped virus 2 (SIRV2) and Sulfolobus turreted icosahedral virus (STIV), are lytic and exploit a special mechanism of virion egress (4–8). During the infection cycle of these viruses, pyramidal protrusions with sevenfold rotational symmetry form in the host cell membrane. As the final step of the infection cycle the virus-associated pyramids (VAPs) open outwards along the seams of their seven facets, creating ∼100-nm apertures through which the newly formed virions escape from the host cell (4, 7). VAPs consist of multiple copies of an ∼10-kDa virus-encoded protein, which we term “PVAP” (Protein forming Virus-Associated Pyramids/SIRV2_P98) (7–9). Surprisingly, PVAP assembles into membrane pyramids even when expressed heterologously in archaeal and bacterial expression systems, demonstrating that no other viral proteins are required for VAP formation (7). The mechanism by which VAPs self-assembles in the membrane remains unknown.In the present study we used electron cryotomography to investigate morphological features of SIRV2 replication and the formation of VAPs at different time points after infection. By subtomogram averaging, we determined a 3D map of the VAP. This map, in combination with secondary structure predictions of PVAP and the expression of wild-type (WT) PVAP or a variety of truncation mutants in archaeal, bacterial and eukaryotic cells allows us to propose a model showing how PVAP arranges to form the sevenfold pyramids. These insights are fundamental for understanding how this mechanism can be exploited as a universal tool to engineer the formation and controlled opening of large pores in biological or artificial lipid bilayers.     

Predicting aggressive behaviour in acute forensic mental health units: A re‐examination of the dynamic appraisal of situational aggression's predictive validity

In the present study, we explored the predictive validity of the Dynamic Appraisal of Situational Aggression (DASA) assessment tool in male (n = 30) and female (n = 30) patients admitted to the acute units of a forensic mental health hospital. We also tested the psychometric properties of the original DASA bands and novel risk bands. The first 60 days of each patient's file was reviewed to identify daily DASA scores and subsequent risk‐related nursing interventions and aggressive behaviour within the following 24 hours. Risk assessments, followed by documented nursing interventions, were removed to preserve the integrity of the risk‐assessment analysis. Receiver–operator characteristics were used to test the predictive accuracy of the DASA, and generalized estimating equations (GEE) were used to account for repeated risk assessments, which occurs when analysing short‐term risk‐assessment data. The results revealed modest predictive validity for males and females. GEE analyses suggested the need to adjust the DASA risk bands to the following (with associated odds ratios (OR) for aggressive behaviour): 0 = low risk; 1, 2, 3 = moderate‐risk OR, 4.70 (95% confidence interval (CI): 2.84–7.80); and 4, 5, 6, 7 = high‐risk OR, 16.13 (95% CI: 9.71–26.78). The adjusted DASA risk bands could assist nurses by prompting violence‐prevention interventions when the level of risk is elevated.     

Linkage disequilibrium in young genetically isolated Dutch population

The design and feasibility of genetic studies of complex diseases are critically dependent on the extent and distribution of linkage disequilibrium (LD) across the genome and between different populations. We have examined genomewide and region-specific LD in a young genetically isolated population identified in the Netherlands by genotyping approximately 800 Short Tandem Repeat markers distributed genomewide across 58 individuals. Several regions were analyzed further using a denser marker map. The permutation-corrected measure of LD was used for analysis. A significant (P<0.0004) relation between LD and genetic distance on a genomewide scale was found. Distance explained 4% of the total LD variation. For fine-mapping data, distance accounted for a larger proportion of LD variation (up to 39%). A notable similarity in the genomewide distribution of LD was revealed between this population and other young genetically isolated populations from Micronesia and Costa Rica. Our study population and experiment was simulated in silico to confirm our knowledge of the history of the population. High agreement was observed between results of analysis of simulated and empirical data. We conclude that our population shows a high level of LD similar to that demonstrated previously in other young genetic isolates. In Europe, there may be a large number of young genetically isolated populations that are similar in history to ours. In these populations, a similar degree of LD is expected and thus they may be effectively used for linkage or LD mapping.     

Alpha waves: a neural signature of visual suppression

Alpha waves are traditionally considered a passive consequence of the lack of stimulation of sensory areas. However, recent results have challenged this view by showing a modulation of alpha activity in cortical areas representing unattended information during active tasks. These data have led us to think that alpha waves would support a 'gating function' on sensorial stimulation that actively inhibits unattended information in attentional tasks. Visual suppression occurring during a saccade and blink entails an inhibition of incoming visual information, and it seems to occur at an early processing stage. In this study, we hypothesized that the neural mechanism through which the visual system exerts this inhibition is the active imposition of alpha oscillations in the occipital cortex, which in turn predicts an increment of alpha amplitude during a visual suppression phenomena. We measured visual suppression occurring during short closures of the eyelids, a situation well suited for EEG recordings and stimulated the retinae with an intra-oral light administered through the palate. In the behavioral experiment, detection thresholds were measured with eyes steady open and steady closed, showing a reduction of sensitivity in the latter case. In the EEG recordings performed under identical conditions we found stronger alpha activity with closed eyes. Since the stimulation does not depend on whether the eyes were open or closed, we reasoned that this should be a central effect, probably due to a functional role of alpha oscillation in agreement with the 'gating function' theory.     

Systolic blood pressure reactions to acute stress are associated with future hypertension status in the Dutch Famine Birth Cohort Study

These analyses examined the association between blood pressure reactions to acute psychological stress and subsequent hypertension status in a substantial Dutch cohort. Blood pressure was recorded during a resting baseline and during three acute stress tasks, Stroop colour word, mirror tracing and speech. Five years later, diagnosed hypertension status was determined by questionnaire. Participants were 453 (237 women) members of the Dutch Famine Birth Cohort. In analysis adjusting for a number of potential confounders, systolic blood pressure reactivity was positively related to future hypertension. This was the case irrespective of whether reactivity was calculated as the peak or the average response to the stress tasks. The association was strongest for reactions to the speech and Stroop tasks. Diastolic blood pressure reactivity was not significantly associated with hypertension. The results provide support for the reactivity hypothesis.     

Novel 7‐DHCR mutation in a child with Smith‐Lemli‐Opitz syndrome

Smith‐Lemli‐Opitz syndrome (SLOS) is an autosomal recessive disorder characterized by minor facial anomalies, mental retardation, and multiple congenital abnormalities. Biochemically, the disorder is caused by deficient activity of 7‐dehydrocholesterol reductase, which catalyzes the reduction of the Δ7 double bond of 7‐dehydrocholesterol to produce cholesterol. Recently, mutations in the gene encoding 7‐dehydrocholesterol reductase (7DHCR) were found to cause SLOS. We report the first molecular characterization of an Italian SLOS patient. Interestingly, his paternal 7DHCR allele, of Arab origin, harbored a novel P329L mutation which in combination with a maternal splice‐site (IVS8‐1 G>C) mutation resulted in a relatively milder phenotype. Am. J. Med. Genet. 91:138–140, 2000. © 2000 Wiley‐Liss, Inc.     

Diminished heart rate reactivity to acute psychological stress is associated with enhanced carotid intima‐media thickness through adverse health behaviors

Recent evidence demonstrates that individuals with low heart rate (HR) reactions to acute psychological stress are more likely to be obese or smokers. Smoking and obesity are established risk factors for increased carotid intima‐media thickness (IMT). The aim of this study was to examine the potential pathways linking intima‐media thickness, smoking, body mass index (BMI), and HR stress reactivity. A total of 552 participants, 47.6% male, M (SD) age = 58.3 (0.94) years, were exposed to three psychological stress tasks (Stroop, mirror drawing, and speech) preceded by a resting baseline period; HR was recorded throughout. HR reactivity was calculated as the average response across the three tasks minus average baseline HR. Smoking status, BMI, and IMT were determined by trained personnel. Controlling for important covariates (e.g., socioeconomic status), structural equation modeling revealed that BMI and smoking mediated the negative relationship between HR reactivity and IMT. The hypothesized model demonstrated a good overall fit to the data, χ²(8) = 0.692, p = .403; CFI = 1.00; TLI = 1.00 SRMR = .01; RMSEA < .001 (90% CI < 0.01–0.11). HR reactivity was negatively related to BMI (β = ?.16) and smoking (β = ?.18), and these in turn were positively associated with IMT (BMI: β = .10; smoking: β = .17). Diminished HR stress reactivity appears to be a marker for enlarged IMT and appears to be exerting its impact through already established risks. Future research should examine this relationship longitudinally and aim to intervene early.