Molecular basis for protection of ribosomal protein L4 from cellular degradation

Eukaryotic ribosome biogenesis requires the nuclear import of ∼80 nascent ribosomal proteins and the elimination of excess amounts by the cellular degradation machinery. Assembly chaperones recognize nascent unassembled ribosomal proteins and transport them together with karyopherins to their nuclear destination. We report the crystal structure of ribosomal protein L4 (RpL4) bound to its dedicated assembly chaperone of L4 (Acl4), revealing extensive interactions sequestering 70 exposed residues of the extended RpL4 loop. The observed molecular recognition fundamentally differs from canonical promiscuous chaperone–substrate interactions. We demonstrate that the eukaryote-specific RpL4 extension harbours overlapping binding sites for Acl4 and the nuclear transport factor Kap104, facilitating its continuous protection from the cellular degradation machinery. Thus, Acl4 serves a dual function to facilitate nuclear import and simultaneously protect unassembled RpL4 from the cellular degradation machinery

Structural and Functional Analysis of Human SIRT1

SIRT1 is a NAD^+-dependent deacetylase that plays important roles in many cellular processes. SIRT1 activity is uniquely controlled by a C-terminal regulatory segment (CTR). Here we present crystal structures of the catalytic domain of human SIRT1 in complex with the CTR in an open apo form and a closed conformation in complex with a cofactor and a pseudo-substrate peptide. The catalytic domain adopts the canonical sirtuin fold. The CTR forms a β hairpin structure that complements the β sheet of the NAD^+-binding domain, covering an essentially invariant hydrophobic surface. The apo form adopts a distinct open conformation, in which the smaller subdomain of SIRT1 undergoes a rotation with respect to the larger NAD^+-binding subdomain. A biochemical analysis identifies key residues in the active site, an inhibitory role for the CTR, and distinct structural features of the CTR that mediate binding and inhibition of the SIRT1 catalytic domain

TERJEMAHAN BERANOTASI DONGENG LE FILS À LA RECHERCHE DE SA MÈRE KE DALAM BAHASA INDONESIA

Annotated translation is a study that provides annotations or notes on the chosen equivalents of a number of translated words as a form of translator’s accountability. Using a comparative model, this qualitative study aims to describe the problems that were encountered when translating the source text and finding the right translation strategy to be used for addressing the existing translation problems. In this research, the source text is a children literature (tale) titled Le Fils à la recherche de sa mère by Senegalese author. The problems that were encountered when translating this tale were issues related to language and culture, such as idioms, metaphors, and cultural words. The translation problems were then addressed by using translation strategies (methods and procedures) according to Newmark (1988). In generating translations and annotations, this research referred to various dictionaries and websites. The findings of this research are expected to enrich the French children literature translations from African countries that are rarely found in Indonesia

Molecular basis for protection of ribosomal protein L4 from cellular degradation

Eukaryotic ribosome biogenesis requires the nuclear import of ∼80 nascent ribosomal proteins and the elimination of excess amounts by the cellular degradation machinery. Assembly chaperones recognize nascent unassembled ribosomal proteins and transport them together with karyopherins to their nuclear destination. We report the crystal structure of ribosomal protein L4 (RpL4) bound to its dedicated assembly chaperone of L4 (Acl4), revealing extensive interactions sequestering 70 exposed residues of the extended RpL4 loop. The observed molecular recognition fundamentally differs from canonical promiscuous chaperone–substrate interactions. We demonstrate that the eukaryote-specific RpL4 extension harbours overlapping binding sites for Acl4 and the nuclear transport factor Kap104, facilitating its continuous protection from the cellular degradation machinery. Thus, Acl4 serves a dual function to facilitate nuclear import and simultaneously protect unassembled RpL4 from the cellular degradation machinery

Histone-Binding of DPF2 Mediates Its Repressive Role in Myeloid Differentiation

Double plant homeodomain finger 2 (DPF2) is a highly evolutionarily conserved member of the d4 protein family that is ubiquitously expressed in human tissues and was recently shown to inhibit the myeloid differentiation of hematopoietic stem/progenitor and acute myelogenous leukemia cells. Here, we present the crystal structure of the tandem plant homeodomain finger domain of human DPF2 at 1.6-Å resolution. We show that DPF2 interacts with the acetylated tails of both histones 3 and 4 via bipartite binding pockets on the DPF2 surface. Blocking these interactions through targeted mutagenesis of DPF2 abolishes its recruitment to target chromatin regions as well as its ability to prevent myeloid differentiation in vivo. Our findings suggest that the histone binding of DPF2 plays an important regulatory role in the transcriptional program that drives myeloid differentiation

Structural and functional analysis of mRNA export regulation by the nuclear pore complex

The nuclear pore complex (NPC) controls the passage of macromolecules between the nucleus and cytoplasm, but how the NPC directly participates in macromolecular transport remains poorly understood. In the final step of mRNA export, the DEAD-box helicase DDX19 is activated by the nucleoporins Gle1, Nup214, and Nup42 to remove Nxf1•Nxt1 from mRNAs. Here, we report crystal structures of Gle1•Nup42 from three organisms that reveal an evolutionarily conserved binding mode. Biochemical reconstitution of the DDX19 ATPase cycle establishes that human DDX19 activation does not require IP_6, unlike its fungal homologs, and that Gle1 stability affects DDX19 activation. Mutations linked to motor neuron diseases cause decreased Gle1 thermostability, implicating nucleoporin misfolding as a disease determinant. Crystal structures of human Gle1•Nup42•DDX19 reveal the structural rearrangements in DDX19 from an auto-inhibited to an RNA-binding competent state. Together, our results provide the foundation for further mechanistic analyses of mRNA export in humans

Focusing a deterministic single-ion beam

We focus down an ion beam consisting of single 40Ca+ ions to a spot size of a few mum using an einzel-lens. Starting from a segmented linear Paul trap, we have implemented a procedure which allows us to deterministically load a predetermined number of ions by using the potential shaping capabilities of our segmented ion trap. For single-ion loading, an efficiency of 96.7(7)% has been achieved. These ions are then deterministically extracted out of the trap and focused down to a 1sigma-spot radius of (4.6 \pm 1.3)mum at a distance of 257mm from the trap center. Compared to former measurements without ion optics, the einzel-lens is focusing down the single-ion beam by a factor of 12. Due to the small beam divergence and narrow velocity distribution of our ion source, chromatic and spherical aberration at the einzel-lens is vastly reduced, presenting a promising starting point for focusing single ions on their way to a substrate.Comment: 16 pages, 7 figure

A conserved quality-control pathway that mediates degradation of unassembled ribosomal proteins

Overproduced yeast ribosomal protein (RP) Rpl26 fails to assemble into ribosomes and is degraded in the nucleus/nucleolus by a ubiquitin-proteasome system quality control pathway comprising the E2 enzymes Ubc4/Ubc5 and the ubiquitin ligase Tom1. tom1 cells show reduced ubiquitination of multiple RPs, exceptional accumulation of detergent-insoluble proteins including multiple RPs, and hypersensitivity to imbalances in production of RPs and rRNA, indicative of a profound perturbation to proteostasis. Tom1 directly ubiquitinates unassembled RPs primarily via residues that are concealed in mature ribosomes. Together, these data point to an important role for Tom1 in normal physiology and prompt us to refer to this pathway as ERISQ, for excess ribosomal protein quality control. A similar pathway, mediated by the Tom1 homolog Huwe1, restricts accumulation of overexpressed hRpl26 in human cells. We propose that ERISQ is a key element of the quality control machinery that sustains protein homeostasis and cellular fitness in eukaryotes

Structural and functional analysis of mRNA export regulation by the nuclear pore complex

The nuclear pore complex (NPC) controls the passage of macromolecules between the nucleus and cytoplasm, but how the NPC directly participates in macromolecular transport remains poorly understood. In the final step of mRNA export, the DEAD-box helicase DDX19 is activated by the nucleoporins Gle1, Nup214, and Nup42 to remove Nxf1•Nxt1 from mRNAs. Here, we report crystal structures of Gle1•Nup42 from three organisms that reveal an evolutionarily conserved binding mode. Biochemical reconstitution of the DDX19 ATPase cycle establishes that human DDX19 activation does not require IP_6, unlike its fungal homologs, and that Gle1 stability affects DDX19 activation. Mutations linked to motor neuron diseases cause decreased Gle1 thermostability, implicating nucleoporin misfolding as a disease determinant. Crystal structures of human Gle1•Nup42•DDX19 reveal the structural rearrangements in DDX19 from an auto-inhibited to an RNA-binding competent state. Together, our results provide the foundation for further mechanistic analyses of mRNA export in humans

Sideband cooling and coherent dynamics in a microchip multi-segmented ion trap

Miniaturized ion trap arrays with many trap segments present a promising architecture for scalable quantum information processing. The miniaturization of segmented linear Paul traps allows partitioning the microtrap in different storage and processing zones. The individual position control of many ions - each of them carrying qubit information in its long-lived electronic levels - by the external trap control voltages is important for the implementation of next generation large-scale quantum algorithms. We present a novel scalable microchip multi-segmented ion trap with two different adjacent zones, one for the storage and another dedicated for the processing of quantum information using single ions and linear ion crystals: A pair of radio-frequency driven electrodes and 62 independently controlled DC electrodes allows shuttling of single ions or linear ion crystals with numerically designed axial potentials at axial and radial trap frequencies of a few MHz. We characterize and optimize the microtrap using sideband spectroscopy on the narrow S1/2 D5/2 qubit transition of the 40Ca+ ion, demonstrate coherent single qubit Rabi rotations and optical cooling methods. We determine the heating rate using sideband cooling measurements to the vibrational ground state which is necessary for subsequent two-qubit quantum logic operations. The applicability for scalable quantum information processing is proven.Comment: 17 pages, 11 figure