N4BP1 restricts HIV-1 and its inactivation by MALT1 promotes viral reactivation.

RNA-modulating factors not only regulate multiple steps of cellular RNA metabolism, but also emerge as key effectors of the immune response against invading viral pathogens including human immunodeficiency virus type-1 (HIV-1). However, the cellular RNA-binding proteins involved in the establishment and maintenance of latent HIV-1 reservoirs have not been extensively studied. Here, we screened a panel of 62 cellular RNA-binding proteins and identified NEDD4-binding protein 1 (N4BP1) as a potent interferon-inducible inhibitor of HIV-1 in primary T cells and macrophages. N4BP1 harbours a prototypical PilT N terminus-like RNase domain and inhibits HIV-1 replication by interacting with and degrading viral mRNA species. Following activation of CD4+ T cells, however, N4BP1 undergoes rapid cleavage at Arg 509 by the paracaspase named mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1). Mutational analyses and knockout studies revealed that MALT1-mediated inactivation of N4BP1 facilitates the reactivation of latent HIV-1 proviruses. Taken together, our findings demonstrate that the RNase N4BP1 is an efficient restriction factor of HIV-1 and suggest that inactivation of N4BP1 by induction of MALT1 activation might facilitate elimination of latent HIV-1 reservoirs.

Morphology and physical-chemical properties of baked nanoporous frustules.

We investigated the morphology and physical-chemical properties of baked and unbaked nanoporous frustules. Scanning electron microscopy (SEM) observations showed that the nanoporous structures of frustules unchanged at 400 degrees C even after baking for 6 h. During baking at 800 degrees C, the frustule structures changed dramatically. On the other hand, Fourier transform infrared spectroscopy (FTIR) of bulk frustule samples indicated that physical-chemical properties of the frustules had clearly changed after baking at not only 800 degrees C but also 400 degrees C. These results showed that the reconstruction of the structures had occurred inside the frustules, even though the morphology of the frustules had not apparently changed at 400 degrees C. In order to characterize the exact shape of the frustules, living diatom cells were grown on a functionalized mica surface, and then baked without any chemical treatment for SEM study. This 'direct baking' technique is effective for comparing minute structures of the frustules, because completed combination of every part of the frustules can be observed.

Diatom cells grown and baked on a functionalized mica surface.

We demonstrate the cultivation of diatom cells on a functionalized mica surface and the preparation of frustules on a mica surface by baking. Diatom cells were successfully grown on a mica surface treated with 3-aminopropyltriethoxysilane. After baking at 400 degrees C for 2 h, frustule structures without the organic components of the diatom cells were successfully observed by scanning electron microscopy and atomic force microscopy. Furthermore, the frustules deformed and became slender when a sample was baked at 800 degrees C for 2 h. Our method is effective for the direct characterization of frustule structures and physical properties without changing the configuration of the diatom cells grown on the mica surface.

Study of the nanoscopic deformation of an annealed nafion film by using atomic force microscopy and a patterned substrate.

We demonstrated the repetitive imaging of the same area of a nafion film before and after annealing by using atomic force microscopy (AFM). In order to find the exact same area of the same sample after changing the cantilever and reattaching the sample, a micropatterned substrate was developed. A micropattern with a 250-500 microm pitch was prepared on the backside of a transparent glass substrate. This pattern includes various signs such as colored letters and numbers at the center of each lattice of the pattern. The nanostructures fabricated by AFM nanolithography on a nafion film using this new method were successfully characterized before and after annealing (over 100 degrees C). The AFM images clearly showed that the nanostructures on a nafion film were dramatically changed by annealing. The data indicated an evidence to understand why the nafion fuel cell does not work well at high temperatures. Our method is probably effective for the study of nanoscopic dynamics in various surface structures.