Characterization of the HIV-1 integrase chromatin- and LEDGF/p75-binding abilities by mutagenic analysis within the catalytic core domain of integrase

<p>Abstract</p> <p>Background</p> <p>During the early stage of HIV-1 replication, integrase (IN) plays important roles at several steps, including reverse transcription, viral DNA nuclear import, targeting viral DNA to host chromatin and integration. Previous studies have demonstrated that HIV-1 IN interacts with a cellular Lens epithelium-derived growth factor (LEDGF/p75) and that this viral/cellular interaction plays an important role for tethering HIV-1 preintegration complexes (PICs) to transcriptionally active units of host chromatin. Meanwhile, other studies have revealed that the efficient knockdown and/or knockout of LEDGF/p75 could not abolish HIV infection, suggesting a LEDGF/p75-independent action of IN for viral DNA chromatin targeting and integration, even though the underlying mechanism(s) is not fully understood.</p> <p>Results</p> <p>In this study, we performed site-directed mutagenic analysis at the C-terminal region of the IN catalytic core domain responsible for IN/chromatin binding and IN/LEDGF/p75 interaction. The results showed that the IN mutations H171A, L172A and EH170,1AA, located in the loop region ₁₇₀EHLK₁₇₃between the α4 and α5 helices of IN, severely impaired the interaction with LEDGF/p75 but were still able to bind chromatin. In addition, our combined knockdown approach for LEDGF/p75 also failed to dissociate IN from chromatin. This suggests that IN has a LEDGF/p75-independent determinant for host chromatin binding. Furthermore, a single-round HIV-1 replication assay showed that the viruses harboring IN mutants capable of LEDGF/p75-independent chromatin binding still sustained a low level of infection, while the chromatin-binding defective mutant was non-infectious.</p> <p>Conclusions</p> <p>All of these data indicate that, even though the presence of LEDGF/p75 is important for a productive HIV-1 replication, IN has the ability to bind chromatin in a LEDGF/p75-independent manner and sustains a low level of HIV-1 infection. Hence, it is interesting to define the mechanism(s) underlying IN-mediated LEDGF/p75-independent chromatin targeting, and further studies in this regard will help for a better understanding of the molecular mechanism of chromatin targeting by IN during HIV-1 infection.</p

Contribution of the C-terminal tri-lysine regions of human immunodeficiency virus type 1 integrase for efficient reverse transcription and viral DNA nuclear import

BACKGROUND: In addition to mediating the integration process, HIV-1 integrase (IN) has also been implicated in different steps during viral life cycle including reverse transcription and viral DNA nuclear import. Although the karyophilic property of HIV-1 IN has been well demonstrated using a variety of experimental approaches, the definition of domain(s) and/or motif(s) within the protein that mediate viral DNA nuclear import and its mechanism are still disputed and controversial. In this study, we performed mutagenic analyses to investigate the contribution of different regions in the C-terminal domain of HIV-1 IN to protein nuclear localization as well as their effects on virus infection. RESULTS: Our analysis showed that replacing lysine residues in two highly conserved tri-lysine regions, which are located within previously described Region C ((235)WKGPAKLLWKGEGAVV) and sequence Q ((211)KELQKQITK) in the C-terminal domain of HIV-1 IN, impaired protein nuclear accumulation, while mutations for RK(263,4 )had no significant effect. Analysis of their effects on viral infection in a VSV-G pseudotyped RT/IN trans-complemented HIV-1 single cycle replication system revealed that all three C-terminal mutant viruses (KK215,9AA, KK240,4AE and RK263,4AA) exhibited more severe defect of induction of β-Gal positive cells and luciferase activity than an IN class 1 mutant D64E in HeLa-CD4-CCR5-β-Gal cells, and in dividing as well as non-dividing C8166 T cells, suggesting that some viral defects are occurring prior to viral integration. Furthermore, by analyzing viral DNA synthesis and the nucleus-associated viral DNA level, the results clearly showed that, although all three C-terminal mutants inhibited viral reverse transcription to different extents, the KK240,4AE mutant exhibited most profound effect on this step, whereas KK215,9AA significantly impaired viral DNA nuclear import. In addition, our analysis could not detect viral DNA integration in each C-terminal mutant infection, even though they displayed various low levels of nucleus-associated viral DNA, suggesting that these C-terminal mutants also impaired viral DNA integration ability. CONCLUSION: All of these results indicate that, in addition to being involved in HIV-1 reverse transcription and integration, the C-terminal tri-lysine regions of IN also contribute to efficient viral DNA nuclear import during the early stage of HIV-1 replication

A New Method for Impeller Inlet Design of Supercritical CO2 Centrifugal Compressors in Brayton Cycles

Supercritical Carbon Dioxide (SCO2) is considered as a potential working fluid in next generation power and energy systems. The SCO2\ua0Brayton cycle is advantaged with higher cycle efficiency, smaller compression work, and more compact layout, as compared with traditional cycles. When the inlet total condition of the compressor approaches the critical point of the working fluid, the cycle efficiency is further enhanced. However, the flow acceleration near the impeller inducer causes the fluid to enter two-phase region, which may lead to additional aerodynamic losses and flow instability. In this study, a new impeller inlet design method is proposed to achieve a better balance among the cycle efficiency, compressor compactness, and inducer condensation. This approach couples a concept of the maximum swallowing capacity of real gas and a new principle for condensation design. Firstly, the mass flow function of real gas centrifugal compressors is analytically expressed by non-dimensional parameters. An optimal inlet flow angle is derived to achieve the maximum swallowing capacity under a certain inlet relative Mach number, which leads to the minimum energy loss and a more compact geometry for the compressor. Secondly, a new condensation design principle is developed by proposing a novel concept of the two-zone inlet total condition for SCO2\ua0compressors. In this new principle, the acceptable acceleration margin (AAM) is derived as a criterion to limit the impeller inlet condensation. The present inlet design method is validated in the design and simulation of a low-flow-coefficient compressor stage based on the real gas model. The mechanisms of flow accelerations in the impeller inducer, which form low-pressure regions and further produce condensation, are analyzed and clarified under different operating conditions. It is found that the proposed method is efficient to limit the condensation in the impeller inducer, keep the compactness of the compressor, and maintain a high cycle efficiency

Prediction of mining-induced surface movement duration based on an improved Knothe time model

The surface movement duration is a key parameter for the evaluation of the stability of surface movement and deformation, and accurately predicting the surface movement duration under different geological conditions and mining intensity is crucial for the stability assessment of the surface and ground facilities. Firstly, a new model assumption is proposed based on the classical Knothe time model assumption, and an improved Knothe time model, which only contains one model parameter and can accurately describe the dynamic subsidence law of the surface, has been developed. Then, based on the improved Knothe time model, a surface movement duration prediction model that can comprehensively consider the mining depth, coal seam thickness and mining speed of the working face is established, and the determination method of model parameters is provided according to the probability integral method. Finally, the accuracy and rationality of the surface movement duration prediction model were verified by using monitoring data of the surface movement duration in 20 mining areas. The results showed that the predicted value of the surface movement duration prediction model based on the improved Knothe time model is highly consistent with the monitoring values in 20 mining areas, and their root mean square error (ERMS) is only 64 days, which is far less than 882 days of the Specifications of Coal Mining and Pillar Retention Under Buildings, Water and Railways and 152 days of the Knothe time model, which verifies the accuracy and rationality of the prediction model of surface movement duration. The surface movement duration is affected by coal seam mining height, average mining depth and coal seam mining speed, and increases nonlinearly with the increase of coal seam mining height, increases linearly with the increase of average mining depth, but decreases nonlinearly with the increase of coal seam mining speed; When the mining speed of coal seam is less than 5 m/d, the surface movement duration can be shortened by appropriately increasing the mining speed of coal seam. The study can provide some reference for the prediction of the stable time of surface movement and deformation in coal mining

Attribute-Based Conditional Proxy Re-Encryption in the Standard Model under LWE

Attribute-based conditional proxy re-encryption (AB-CPRE) allows delegators to carry out attribute-based control on the delegation of decryption by setting policies and attribute vectors. The fine-grained control of AB-CPRE makes it suitable for a variety of applications, such as cloud storage and distributed file systems. However, all existing AB-CPRE schemes are constructed under classical number-theoretic assumptions, which are vulnerable to quantum cryptoanalysis. Therefore, we propose the first AB-CPRE scheme based on the learning with errors (LWE) assumption. Constructed from fully key-homomorphic encryption (FKHE) and key-switching techniques, our scheme is unidirectional, single-hop, and enables a polynomial-deep boolean circuit as its policy. Furthermore, we split the ciphertext into two independent parts to avoid two-level or multi-level encryption/decryption mechanisms. Taking advantage of it, we then extend our single-hop AB-CPRE into an efficient and concise multi-hop one. No matter how many transformations are performed, the re-encrypted ciphertext is in constant size, and only one encryption/decryption algorithm is needed. Both of our schemes are proved to be selective secure against chosen-plaintext attacks (CPA) in the standard model

Contribution of the C-terminal region within the catalytic core domain of HIV-1 integrase to yeast lethality, chromatin binding and viral replication

<p>Abstract</p> <p>Background</p> <p>HIV-1 integrase (IN) is a key viral enzymatic molecule required for the integration of the viral cDNA into the genome. Additionally, HIV-1 IN has been shown to play important roles in several other steps during the viral life cycle, including reverse transcription, nuclear import and chromatin targeting. Interestingly, previous studies have demonstrated that the expression of HIV-1 IN induces the lethal phenotype in some strains of <it>Saccharomyces cerevisiae</it>. In this study, we performed mutagenic analyses of the C-terminal region of the catalytic core domain of HIV-1 IN in order to delineate the critical amino acid(s) and/or motif(s) required for the induction of the lethal phenotype in the yeast strain HP16, and to further elucidate the molecular mechanism which causes this phenotype.</p> <p>Results</p> <p>Our study identified three HIV-1 IN mutants, V165A, A179P and KR186,7AA, located in the C-terminal region of the catalytic core domain of IN that do not induce the lethal phenotype in yeast. Chromatin binding assays in yeast and mammalian cells demonstrated that these IN mutants were impaired for the ability to bind chromatin. Additionally, we determined that while these IN mutants failed to interact with LEDGF/p75, they retained the ability to bind Integrase interactor 1. Furthermore, we observed that VSV-G-pseudotyped HIV-1 containing these IN mutants was unable to replicate in the C8166 T cell line and this defect was partially rescued by complementation with the catalytically inactive D64E IN mutant.</p> <p>Conclusion</p> <p>Overall, this study demonstrates that three mutations located in the C-terminal region of the catalytic core domain of HIV-1 IN inhibit the IN-induced lethal phenotype in yeast by inhibiting the binding of IN to the host chromatin. These results demonstrate that the C-terminal region of the catalytic core domain of HIV-1 IN is important for binding to host chromatin and is crucial for both viral replication and the promotion of the IN-induced lethal phenotype in yeast.</p

High atmospheric carbon dioxide-dependent alleviation of salt stress is linked to RESPIRATORY BURST OXIDASE 1 (RBOH1)-dependent H2O2 production in tomato (Solanum lycopersicum)

Plants acclimate rapidly to stressful environmental conditions. Increasing atmospheric CO2 levels are predicted to influence tolerance to stresses such as soil salinity but the mechanisms are poorly understood. To resolve this issue, tomato (Solanum lycopersicum) plants were grown under ambient (380 μmol mol–1) or high (760 μmol mol–1) CO2 in the absence or presence of sodium chloride (100 mM). The higher atmospheric CO2 level induced the expression of RESPIRATORY BURST OXIDASE 1 (SlRBOH1) and enhanced H2O2 accumulation in the vascular cells of roots, stems, leaf petioles, and the leaf apoplast. Plants grown with higher CO2 levels showed improved salt tolerance, together with decreased leaf transpiration rates and lower sodium concentrations in the xylem sap, vascular tissues, and leaves. Silencing SlRBOH1 abolished high CO2 -induced salt tolerance and increased leaf transpiration rates, as well as enhancing Na+ accumulation in the plants. The higher atmospheric CO2 level increased the abundance of a subset of transcripts involved in Na+ homeostasis in the controls but not in the SlRBOH1-silenced plants. It is concluded that high atmospheric CO2 concentrations increase salt stress tolerance in an apoplastic H2O2 dependent manner, by suppressing transpiration and hence Na+ delivery from the roots to the shoots, leading to decreased leaf Na+ accumulation

Continuous spin excitations in the three-dimensional frustrated magnet K2Ni2(SO4)3

Continuous spin excitations are widely recognized as one of the hallmarks of novel spin states in quantum magnets, such as quantum spin liquids (QSLs). Here, we report the observation of such kind of excitations in K2Ni2(SO4)3, which consists of two sets of intersected spin-1 Ni2+ trillium lattices. Our inelastic neutron scattering measurement on single crystals clearly shows a dominant excitation continuum, which exhibits a distinct temperature-dependent behavior from that of spin waves, and is rooted in strong quantum spin fluctuations. Further using the self-consistent-gaussian-approximation method, we determined the fourth- and fifth-nearest neighbor exchange interactions are dominant. These two bonds together form a unique three-dimensional network of corner-sharing tetrahedra, which we name as ''hyper-trillium'' lattice. Our results provide direct evidence for the existence of QSL features in K2Ni2(SO4)3 and highlight the potential for the hyper-trillium lattice to host frustrated quantum magnetism.Comment: 6 pages and 5 figures, plus several pages of supplemental material, comments are welcom

Sphingomyelinase inhibitory and free radical scavenging potential of selected Nigerian medicinal plant extracts

Ceramides from sphingolipid breakdown, and other sphingolipid metabolites, mediate cellular signalling in infectious and other diseases. Therefore, inhibitors of sphingomyelinases (SMases), hold promise as prospective therapeutic agents. Considering the potential therapeutic utility, this in vitro study explored the sphingomyelinase inhibitory, and free radical scavenging potential of five Nigerian medicinal plant leaf extracts, purported to have efficacy against diseases, including HIV/AIDS. The extracts\u2019 sphingomyelinase inhibitory potencies were assessed colorimetrically and theirfree radical scavenging capabilities were assayed by the ability to quench 2,2\u2010diphenyl\u20101\u2010picrylhydrazyl (DPPH) radical and superoxide anion (O2.\u2010) radical. Considering their IC50 (\u3bcg/ml) values, the extracts inhibited the biochemical activity of sphingomyelinase in a dose-dependent manner, relative to imipramine the standard inhibitor (IC50 38.5 \ub1 2.4 \u3bcg/ml). With Aloe vera as least inhibitory, inhibition increased as follows: Aloe vera (Asphodelaceae) (1132 \ub1 10.8) &lt; Senna siamea (Fabaceae) (992.2 \ub1 11.2) &lt; Azadirachta indica (Meliaceae) (984 \ub1 7.4) &lt; Landolphia owariensis (Apocynaceae) (146.3 \ub1 9.4) &lt; Stachytarpheta angustifolia (Verbenacae) (100.3 \ub1 8.7). DPPH radical scavenging relative to ascorbic acid standard increased as: A. indica &lt; A. vera &lt; S. siamea &lt; S. angustifolia &lt; L. owariensis; and superoxide anion quenching, relative to standard rutin increased as: A. vera &lt; S. angustifolia &lt; L. owariensis &lt; S. siamea &lt; A. indica.These results showed thatthe most potent SMase inhibitor was S. angustifolia; whereas, for DPPH radical scavenging and superoxide inhibition, the most potent of the five extracts were L. owariensis and A. indica respectively.These extracts deserve further investigation into their biological effects