Binding to DCAF1 distinguishes TASOR and SAMHD1 degradation by HIV-2 Vpx.

Human Immunodeficiency viruses type 1 and 2 (HIV-1 and HIV-2) succeed to evade host immune defenses by using their viral auxiliary proteins to antagonize host restriction factors. HIV-2/SIVsmm Vpx is known for degrading SAMHD1, a factor impeding the reverse transcription. More recently, Vpx was also shown to counteract HUSH, a complex constituted of TASOR, MPP8 and periphilin, which blocks viral expression from the integrated viral DNA. In a classical ubiquitin ligase hijacking model, Vpx bridges the DCAF1 ubiquitin ligase substrate adaptor to SAMHD1, for subsequent ubiquitination and degradation. Here, we investigated whether the same mechanism is at stake for Vpx-mediated HUSH degradation. While we confirm that Vpx bridges SAMHD1 to DCAF1, we show that TASOR can interact with DCAF1 in the absence of Vpx. Nonetheless, this association was stabilized in the presence of Vpx, suggesting the existence of a ternary complex. The N-terminal PARP-like domain of TASOR is involved in DCAF1 binding, but not in Vpx binding. We also characterized a series of HIV-2 Vpx point mutants impaired in TASOR degradation, while still degrading SAMHD1. Vpx mutants ability to degrade TASOR correlated with their capacity to enhance HIV-1 minigenome expression as expected. Strikingly, several Vpx mutants impaired for TASOR degradation, but not for SAMHD1 degradation, had a reduced binding affinity for DCAF1, but not for TASOR. In macrophages, Vpx R34A-R42A and Vpx R42A-Q47A-V48A, strongly impaired in DCAF1, but not in TASOR binding, could not degrade TASOR, while being efficient in degrading SAMHD1. Altogether, our results highlight the central role of a robust Vpx-DCAF1 association to trigger TASOR degradation. We then propose a model in which Vpx interacts with both TASOR and DCAF1 to stabilize a TASOR-DCAF1 complex. Furthermore, our work identifies Vpx mutants enabling the study of HUSH restriction independently from SAMHD1 restriction in primary myeloid cells.

HIV-2/SIV viral protein X counteracts HUSH repressor complex.

To evade host immune defences, human immunodeficiency viruses 1 and 2 (HIV-1 and HIV-2) have evolved auxiliary proteins that target cell restriction factors. Viral protein X (Vpx) from the HIV-2/SIVsmm lineage enhances viral infection by antagonizing SAMHD1 (refs 1,2), but this antagonism is not sufficient to explain all Vpx phenotypes. Here, through a proteomic screen, we identified another Vpx target-HUSH (TASOR, MPP8 and periphilin)-a complex involved in position-effect variegation3. HUSH downregulation by Vpx is observed in primary cells and HIV-2-infected cells. Vpx binds HUSH and induces its proteasomal degradation through the recruitment of the DCAF1 ubiquitin ligase adaptor, independently from SAMHD1 antagonism. As a consequence, Vpx is able to reactivate HIV latent proviruses, unlike Vpx mutants, which are unable to induce HUSH degradation. Although antagonism of human HUSH is not conserved among all lentiviral lineages including HIV-1, it is a feature of viral protein R (Vpr) from simian immunodeficiency viruses (SIVs) of African green monkeys and from the divergent SIV of l'Hoest's monkey, arguing in favour of an ancient lentiviral species-specific vpx/vpr gene function. Altogether, our results suggest the HUSH complex as a restriction factor, active in primary CD4+ T cells and counteracted by Vpx, therefore providing a molecular link between intrinsic immunity and epigenetic control.

Controlled GPR grave research: comparison of reflection profiles between 500 and 250 MHz antennae.

Since ground-penetrating radar (GPR) has become a popular search option for clandestine graves, controlled research is essential to determine the numerous variables that affect grave detection. The purpose of this study was to compare GPR reflection profiles of a controlled grave containing a large pig carcass and a blank control grave at 6 months interment in a Spodosol, which is a common soil type in Florida. Data collection was performed in perpendicular orientations over the graves using both 500 and 250 MHz antennae. Since reflection profiles are used to make initial in-field assessments during a forensic search, it is important for controlled research to evaluate this imagery option. Overall, it was possible to detect the grave containing a pig carcass at 6 months interment that was buried in a Spodosol using both the 500 and the 250 MHz antennae. While the 500 MHz antenna provided more detail within the grave containing a pig carcass, including detecting a soil disturbance and the pig carcass, the 250 MHz antenna also provided excellent imagery. Either antenna would provide optimal results for the type of soil that was sampled. Furthermore, it may be possible to locate actual forensic graves in this soil type when no response from the body is noted, as there may be a discernable response from the disturbed soil within the grave shaft and a noticeable disruption of the spodic horizon. Finally, survey orientation may also affect detection. Since data collection performed in two perpendicular directions detected the pig carcass and the grave floor of the control grave, data collection for an actual search involving a body interred for a long postmortem interval should be performed in both directions when time permits.

The effect of dietary nicotine on the allocation of assimilated food to energy metabolism and growth in fourth-instar larvae of the southern armyworm, Spodoptera eridania (Lepidoptera: Noctuidae).

Dietary nicotine (0.5%), which is a substrate of the PSMO (polysubstrate monooxygenase) detoxification system in the southern armyworm Spodoptera eridania, has significant negative effects on the weight of food ingested, weight gained, relative growth rate (RGR), and efficiency of conversion of digested food (ECD) by fourthinstar S. eridania larvae on a nutrient-rich artificial diet. It has a significant positive effect on the weight of food respired by the larvae. Thus, the detoxification of nicotine by the PSMO system exacts a fitness cost and imposes a metabolic cost on S. eridania larvae. In contrast, dietary α-(+)-pinene, an inducer of the PSMO system, neither exacts a fitness cost nor imposes a metabolic cost on the larvae. We believe this to be the first study to demonstrate unequivocally that the negative effect of a dietary toxin on net growth efficiency (ECD) in an insect herbivore is due to an increase in the allocation of assimilated food to energy metabolism and not to a decrease in the amount of food assimilated. This study, therefore, supports the hypothesis that detoxification can impose a significant metabolic load on an insect herbivore. Implications of a corroboration of the metabolic load hypothesis are discussed.

The effect of acquired microbial enzymes on assimilation efficiency in the common woodlouse, Tracheoniscus rathkei.

The digestive tract of the common woodlouse, Tracheoniscus rathkei Brandt (Isopoda: Oniscoidea), contains digestive enzymes active against α-1,4-glucans, which are the chief storage polysaccharides of vascular plants, algae, fungi, and animals, and ß-1,3-glucans, which are present in algae and fungi. Digestive tract extracts also exhibit significant activity toward xylan and carboxymethyl-cellulose but negligible activity toward microcrystalline cellulose, substrates representative of the major structural polysaccharides of vascular plants. Low activity was detected toward pectin, and no activity was detected toward chitin. Activity toward xylan is due in part to microbial enzymes acquired from the leaf litter which was the isopod's normal food. Although ingested microbial xylanases are stable and active in the gut fluid, they do not make a quantitatively significant contribution to the isopod's ability to assimilate the hemicellulosic component of its diet. However, the assimilation of carbon from labeled plant fiber is enhanced in isopods which have acquired a cellulase by ingestion of leaf litter amended with a commercial preparation of the cellulase complex from the fungus, Penicillium funiculosum. This result demonstrates the potential contribution of acquired enzymes to the digestion of plant fiber in terrestrial detritivores. We urge caution, however, in assigning an important digestive function to ingested enzymes on the basis of evidence that only indicates that such enzymes are present in the gut fluid without additional evidence that their presence results in an enhancement of digestive efficiency.

The transformation of Saperda calcarata (Coleoptera: Cerambycidae) into a cellulose digester through the inclusion of fungal enzymes in its diet.

The larvae of the aspen borer, Saperda calcarata, which feed on the inner bark and sapwood of living aspen stems, are unable to digest cellulose. However, they can be transformed into cellulose digesters by adding the active cellulase complex of the fungus, Penicillium funiculosum to their diet. S. calcarata larvae are preadapted to exploit the digestive potential of ingested microbial enzymes. We argue that ingested fungal enzymes may be responsible for cellulose digestion in many, perhaps most or even all, cellulose digesting cerambycid beetles.

Surfactants: their role in preventing the precipitation of proteins by tannins in insect guts.

Much more tannic acid or pin oak tannin is required to precipitate the abundant leaf protein, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPC), from Manduca sexta gut fluid adjusted to pH 6.5 than is required to precipitate this protein from an aqueous buffer at the same pH. This finding demonstrates that some characteristic of M. sexta gut fluid, in addition to its basicity, counteracts the potential of tannins to precipitate ingested proteins. Gut fluid of M. sexta has a surface tension of 36-39 dynes/cm, indicating the presence of surfactants. Lysolecithin and linoleoylglycine, surfactants known to be present in insect gut fluids, also interfere with the precipitation of RuBPC by tannins at pH 6.5. It is concluded that detergency is a widespread property of insect gut fluids that counteracts the potential of tannins to precipitate die ary proteins, and it is argued that there is no longer any justification for continuing to refer to tannins as digestibility-reducing-substances. Finding that there has been no formidable barrier to the evolution of mechanisms that counter a generalized antidigestive action by tannins is difficult to reconcile with the idea that reduced digestibility is an evolved anti-herbivore adaptation of apparent plants.

Tannin assays in ecological studies: Lack of correlation between phenolics, proanthocyanidins and protein-precipitating constituents in mature foliage of six oak species.

There is no correlation between protein-precipitating capacity and either total phenolic or proanthocyanidin content of extracts of mature foliage from six species of oaks: Quercus alba (white oak), Q. bicolor (swamp white oak), Q. macrocarpa (bur oak), Q. palustris (pin oak), Q. rubra (red oak), and Q. velutina (black oak). It is argued that studies which probe the role of tannins in the selection and utilization of food by herbivores should include a protein-precipitation assay, since such an assay provides a measure of the property of tannins which is presumed to contribute to their utility as defensive compounds. A convenient modification of the bovine serum albumin (BSA) precipitation assay, which measures the amount of protein precipitated when a plant extract is added to a BSA solution, is described. Advantages of this procedure recommend its routine adoption in studies of the role of tannins in plant-herbivore interactions.