Biochemical and virological analysis of the 18-residue C-terminal tail of HIV-1 integrase

Background The 18 residue tail abutting the SH3 fold that comprises the heart of the C-terminal domain is the only part of HIV-1 integrase yet to be visualized by structural biology. To ascertain the role of the tail region in integrase function and HIV-1 replication, a set of deletion mutants that successively lacked three amino acids was constructed and analyzed in a variety of biochemical and virus infection assays. HIV-1/2 chimers, which harbored the analogous 23-mer HIV-2 tail in place of the HIV-1 sequence, were also studied. Because integrase mutations can affect steps in the replication cycle other than integration, defective mutant viruses were tested for integrase protein content and reverse transcription in addition to integration. The F185K core domain mutation, which increases integrase protein solubility, was furthermore analyzed in a subset of mutants. Results Purified proteins were assessed for in vitro levels of 3' processing and DNA strand transfer activities whereas HIV-1 infectivity was measured using luciferase reporter viruses. Deletions lacking up to 9 amino acids (1-285, 1-282, and 1-279) displayed near wild-type activities in vitro and during infection. Further deletion yielded two viruses, HIV-11-276 and HIV-11-273, that displayed approximately two and 5-fold infectivity defects, respectively, due to reduced integrase function. Deletion mutant HIV-11-270 and the HIV-1/2 chimera were non-infectious and displayed approximately 3 to 4-fold reverse transcription in addition to severe integration defects. Removal of four additional residues, which encompassed the C-terminal β strand of the SH3 fold, further compromised integrase incorporation into virions and reverse transcription. Conclusion HIV-11-270, HIV-11-266, and the HIV-1/2 chimera were typed as class II mutant viruses due to their pleiotropic replication defects. We speculate that residues 271-273 might play a role in mediating the known integrase-reverse transcriptase interaction, as their removal unveiled a reverse transcription defect. The F185K mutation reduced the in vitro activities of 1-279 and 1-276 integrases by about 25%. Mutant proteins 1-279/F185K and 1-276/F185K are therefore highlighted as potential structural biology candidates, whereas further deleted tail variants (1-273/F185K or 1-270/F185K) are less desirable due to marginal or undetectable levels of integrase function

A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors

Host proteins are essential for HIV entry and replication and can be important nonviral therapeutic targets. Large-scale RNA interference (RNAi)-based screens have identified nearly a thousand candidate host factors, but there is little agreement among studies and few factors have been validated. Here we demonstrate that a genome-wide CRISPR-based screen identifies host factors in a physiologically relevant cell system. We identify five factors, including the HIV co-receptors CD4 and CCR5, that are required for HIV infection yet are dispensable for cellular proliferation and viability. Tyrosylprotein sulfotransferase 2 (TPST2) and solute carrier family 35 member B2 (SLC35B2) function in a common pathway to sulfate CCR5 on extracellular tyrosine residues, facilitating CCR5 recognition by the HIV envelope. Activated leukocyte cell adhesion molecule (ALCAM) mediates cell aggregation, which is required for cell-to-cell HIV transmission. We validated these pathways in primary human CD4 + T cells through Cas9-mediated knockout and antibody blockade. Our findings indicate that HIV infection and replication rely on a limited set of host-dispensable genes and suggest that these pathways can be studied for therapeutic intervention

Pengembangan Game Platformer 2D Menggunakan Teknik Projection Mapping

Augmented reality merupakan salah satu trend yang baru di dalam bermain video game dan populer di kalangan pecinta video game sekarang ini. Augmented reality menjadi populer di kalangan pecinta game khususnya disebabkan tingkat interaktif dan immersive yang sangat tinggi pada game yang menggunakan teknik augmented reality tesebut. Salah satu teknik augmented reality yang dapat digunakan dalam bermain game adalah dengan menggunakan teknik projection mapping. Oleh karena itu, dalam proyek skripsi ini, dibangun sebuah game platformer 2D menggunakan teknik projection mapping. Game ini dirancang dengan menggunakan metode formal elements dalam game, dan selanjutnya menggunakan digital prototype dalam mengembangkan prototype game ini. Berdasarkan hasil pengujian fungsionalitas dengan menggunakan basis path testing maka ditunjukkan bahwa fungsionalitas game platformer 2D “Slime Collector” telah berjalan dengan baik. Berdasarkan hasil pengujian gameplay testing dapat disimpulkan bahwa game platformer 2D yang telah dikembangkan dengan teknik projection mapping ini cukup menarik, interaktif, dan immersive

Assemblage des particules virales et transmission du VIH de cellule à cellule

PARIS7-Bibliothèque centrale (751132105) / SudocSudocFranceF

La chirurgie de l'obésité vue par les médecins généralistes (enquête auprès d'un panel de médecins généralistes de l'Hérault)

MONTPELLIER-BU Médecine UPM (341722108) / SudocMONTPELLIER-BU Médecine (341722104) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

Effect of Superficial Atmospheric Corrosion Upon the Internal Stresses in Structural Steel Elements

A research program is presented showing the stress status determined by the corrosion phenomenon inside a specimen of a structural steel element. Several stains are studied their diameters ranging from 1~mm to 6~mm and thickness of the corroded layer under 0.5~mm. The physical modeling is the result of testing in laboratory the phenomenon of superficial atmospheric corrosion and the numerical modeling was developed under a FEM program, ALGOR. A number of 3,200 finite elements of BRICK type were created and the evolution of normal and tangential stresses was scrutinized under the process of loosing elementary material transformed into scrap. Stresses in the damaged sphere were graphically put into evidence and determined with accuracy due to the performances of the program, showing the local perturbations and the pattern of stress concentrators. The studies showed the importance of reproducing with both physical and mathematical methods the intricate mechanism and sometimes unpredictable effects of corrosion phenomenon upon the structural steel elements

Por los herederos del Contador Iuan Lopez de Ibartola con doña Susana Monel, hija, y heredera de doña Sarra Monel ..

A 109/136(19