Dual role of TRBP in HIV replication and RNA interference: viral diversion of a cellular pathway or evasion from antiviral immunity?

Increasing evidence indicates that RNA interference (RNAi) may be used to provide antiviral immunity in mammalian cells. Human micro (mi)RNAs can inhibit the replication of a primate virus, whereas a virally-encoded miRNA from HIV inhibits its own replication. Indirect proof comes from RNAi suppressors encoded by mammalian viruses. Influenza NS1 and Vaccinia E3L proteins can inhibit RNAi in plants, insects and worms. HIV-1 Tat protein and Adenovirus VA RNAs act as RNAi suppressors in mammalian cells. Surprisingly, many RNAi suppressors are also inhibitors of the interferon (IFN)-induced protein kinase R (PKR) but the potential overlap between the RNAi and the IFN pathways remains to be determined. The link between RNAi as an immune response and the IFN pathway may be formed by a cellular protein, TRBP, which has a dual role in HIV replication and RNAi. TRBP has been isolated as an HIV-1 TAR RNA binding protein that increases HIV expression and replication by inhibiting PKR and by increasing translation of structured RNAs. A recent report published in the Journal of Virology shows that the poor replication of HIV in astrocytes is mainly due to a heightened PKR response that can be overcome by supplying TRBP exogenously. In two recent papers published in Nature and EMBO Reports, TRBP is now shown to interact with Dicer and to be required for RNAi mediated by small interfering (si) and micro (mi)RNAs. The apparent discrepancy between TRBP requirement in RNAi and in HIV replication opens the hypotheses that RNAi may be beneficial for HIV-1 replication or that HIV-1 may evade the RNAi restriction by diverting TRBP from Dicer and use it for its own benefit

Stereoselective synthesis of fluorinated galactopyranosides as potential molecular probes for galactophilic proteins : assessment of monofluorogalactoside–LecA interactions

The replacement of hydroxyl groups by fluorine atoms on hexopyranoside scaffolds may allow access to invaluable tools for studying various biochemical processes. As part of ongoing activities toward the preparation of fluorinated carbohydrates, a systematic investigation involving the synthesis and biological evaluation of a series of mono‐ and polyfluorinated galactopyranosides is described. Various monofluorogalactopyranosides, a trifluorinated, and a tetrafluorinated galactopyranoside have been prepared using a Chiron approach. Given the scarcity of these compounds in the literature, in addition to their synthesis, their biological profiles were evaluated. Firstly, the fluorinated compounds were investigated as antiproliferative agents using normal human and mouse cells in comparison with cancerous cells. Most of the fluorinated compounds showed no antiproliferative activity. Secondly, these carbohydrate probes were used as potential inhibitors of galactophilic lectins. The first transverse relaxation‐optimized spectroscopy (TROSY) NMR experiments were performed on these interactions, examining chemical shift perturbations of the backbone resonances of LecA, a virulence factor from Pseudomonas aeruginosa. Moreover, taking advantage of the fluorine atom, the 19F NMR resonances of the monofluorogalactopyranosides were directly monitored in the presence and absence of LecA to assess ligand binding. Lastly, these results were corroborated with the binding potencies of the monofluorinated galactopyranoside derivatives by isothermal titration calorimetry experiments. Analogues with fluorine atoms at C‐3 and C‐4 showed weaker affinities with LecA as compared to those with the fluorine atom at C‐2 or C‐6. This research has focused on the chemical synthesis of “drug‐like” low‐molecular‐weight inhibitors that circumvent drawbacks typically associated with natural oligosaccharides

Characterization of the TRBP domain required for Dicer interaction and function in RNA interference

<p>Abstract</p> <p>Background</p> <p>Dicer, Ago2 and TRBP are the minimum components of the human RNA-induced silencing complex (RISC). While Dicer and Ago2 are RNases, TRBP is the double-stranded RNA binding protein (dsRBP) that loads small interfering RNA into the RISC. TRBP binds directly to Dicer through its C-terminal domain.</p> <p>Results</p> <p>We show that the TRBP binding site in Dicer is a 165 amino acid (aa) region located between the ATPase and the helicase domains. The binding site in TRBP is a 69 aa domain, called C4, located at the C-terminal end of TRBP. The TRBP1 and TRBP2 isoforms, but not TRBPs lacking the C4 site (TRBPsΔC4), co-immunoprecipitated with Dicer. The C4 domain is therefore necessary to bind Dicer, irrespective of the presence of RNA. Immunofluorescence shows that while full-length TRBPs colocalize with Dicer, TRBPsΔC4 do not. <it>tarbp2</it>^-/-cells, which do not express TRBP, do not support RNA interference (RNAi) mediated by short hairpin or micro RNAs against EGFP. Both TRBPs, but not TRBPsΔC4, were able to rescue RNAi function. In human cells with low RNAi activity, addition of TRBP1 or 2, but not TRBPsΔC4, rescued RNAi function.</p> <p>Conclusion</p> <p>The mapping of the interaction sites between TRBP and Dicer show unique domains that are required for their binding. Since TRBPsΔC4 do not interact or colocalize with Dicer, we suggest that TRBP and Dicer, both dsRBPs, do not interact through bound dsRNA. TRBPs, but not TRBPsΔC4, rescue RNAi activity in RNAi-compromised cells, indicating that the binding of Dicer to TRBP is critical for RNAi function.</p

Expression of viral polymerase and phosphorylation of core protein determine core and capsid localization of the human hepatitis B virus

Biopsies from patients show that hepadnaviral core proteins and capsids-collectively called core-are found in the nucleus and cytoplasm of infected hepatocytes. In the majority of studies, cytoplasmic core localization is related to low viraemia while nuclear core localization is associated with high viral loads. In order to better understand the molecular interactions leading to core localization, we analysed transfected hepatoma cells using immune fluorescence microscopy. We observed that expression of core protein in the absence of other viral proteins led to nuclear localization of core protein and capsids, while expression of core in the context of the other viral proteins resulted in a predominantly cytoplasmic localization. Analysis of which viral partner was responsible for cytoplasmic retention indicated that the HBx, surface proteins and HBeAg had no impact but that the viral polymerase was the major determinant. Further analysis revealed that e, an RNA structure to which the viral polymerase binds, was essential for cytoplasmic retention. Furthermore, we showed that core protein phosphorylation at Ser 164 was essential for the cytoplasmic core localization phenotype, which is likely to explain differences observed between individual cells

Production of H2 by water radiolysis in cement paste under electron irradiation: A joint experimental and theoretical study

International audienceLong-term confinement of nuclear waste is one of the main challenges faced by the nuclear industry. Fission products such as 90 Sr and 137 Cs, both β − emitters known to induce serious health hazards, represent the largest fraction of nuclear waste. Cement is a good candidate to store them, provided it can resist the effects of irradiation over time. Here, we have investigated the effects of β − decay on cement by performing electron irradiation experiments on different samples. We show that H 2 production in cement, the main effect of water radiolysis, depends strongly on composition and relative humidity. First-principles calculations indicate that the water-rich interlayer regions with Ca 2+ ions act as electron traps that promote the formation of H 2. They also show that holes localize in water-rich regions in low Ca content samples and are then able to participate in H 2 production. This work provides new understanding of radiolysis effects in cements

Nature(s) au travail

Ce numéro de la Revue d’Anthropologie des Connaissances propose une réflexion sur le développement de propositions scientifiques dans les sciences humaines et sociales à propos du « travail de la nature »

Naturaleza(s) trabajando. Qué hacen los estudios animales y ambientales con el trabajo

International audienceEste número de la Revue d’Anthropologie des Connaissances plantea una reflexión sobre el desarrollo de proposiciones científicas dentro de las ciencias humanas y sociales a propósito del «trabajo de la naturaleza». La presente introducción elabora una cartografía general, mas no exhaustiva, de los usos de la noción de trabajo en los regímenes de producción de conocimientos de los estudios animales y ambientales. Se demuestra primero la dimensión pionera de los estudios animales, cuyos trabajos fundantes pusieron en tela de juicio el legado marxista en cuanto a la relación entre trabajo, subjetividad y valores. Tales trabajos alimentaron luego la reflexión en los campos de la ética y la economía política. Esta cartografía, además, echa luz sobre el desplazamiento de las fronteras del trabajo más allá del animal en las humanidades ambientales. Se plasma la influencia de los estudios animales, así como las particularidades y controversias de las aproximaciones de las humanidades ambientales, en su manera de revisar la separación naturaleza-trabajo para responder a la cuestión ecológica. Por último, se interroga el entusiasmo por el trabajo en los estudios animales y medioambientales como modo de conocer y modificar nuestra relación con los animales y la naturaleza

Nature(s) au travail. Ce que font les études animales et environnementales avec le travail

International audienceCe numéro de la Revue d’Anthropologie des Connaissances propose une réflexion sur le développement de propositions scientifiques dans les sciences humaines et sociales à propos du « travail de la nature ». Cette introduction dresse une cartographie générale, mais non exhaustive, des usages de la notion de travail dans les régimes de production de connaissances des études animales et environnementales. Elle montre d’abord la dimension pionnière des études animales, où des travaux fondateurs ont remis en question des legs marxistes sur les rapports entre travail, subjectivité et valeurs. Ces travaux ont alimenté par la suite des réflexions dans le champ de l’éthique et de l’économie politique. Cette cartographie met ensuite en lumière le déplacement des frontières du travail au-delà de l’animal dans les humanités environnementales. Elle montre l’influence des études animales, ainsi que les particularités et controverses des approches des humanités environnementales dans leur manière de réviser la séparation nature-travail pour répondre à la question écologique. Cette cartographie interroge l’engouement pour le travail dans les études animales et environnementales, comme manière de connaître et de changer nos rapports aux animaux et à la nature