Beta-Detected NQR in Zero Field with a Low Energy Beam of \nuc{8}{Li}+^+

Beta-detected nuclear quadrupole resonances (\bnqr) at zero field are observed using a beam of low energy highly polarized radioactive \nuc{8}{Li}$^+$. The resonances were detected in SrTiO$_3$, Al$_2$O$_3$ and Sr$_2$RuO$_4$ single crystals by monitoring the beta-decay anisotropy as a function of a small audio frequency magnetic field. The resonances show clearly that \nuc{8}{Li} occupies one site with non-cubic symmetry in SrTiO$_3, two in Al$_2$O$_3$and three sites in Sr$_2$RuO$_4$. The resonance amplitude and width are surprisingly large compared to the values expected from transitions between the$|\pm 2> \leftrightarrow |\pm 1>$spin states, indicating a significant mixing between the$|\pm m>$ quadrupolar split levels.Comment: accepted for publication in Physica

Trypanosomatid RACK1 Orthologs Show Functional Differences Associated with Translation Despite Similar Roles in Leishmania Pathogenesis

RACK1 proteins belong to the eukaryote WD40-repeat protein family and function as spatial regulators of multiple cellular events, including signaling pathways, the cell cycle and translation. For this latter role, structural and genetic studies indicate that RACK1 associates with the ribosome through two conserved positively charged amino acids in its first WD40 domain. Unlike RACK1s, including Trypanosoma brucei RACK1 (TbRACK1), only one of these two positively-charged residues is conserved in the first WD40 domain of the Leishmania major RACK1 ortholog, LACK. We compared virulence-attenuated LACK single copy (LACK/-) L. major, with L. major expressing either two LACK copies (LACK/LACK), or one copy each of LACK and TbRACK1 (LACK/TbRACK1), to evaluate the function of these structurally distinct RACK1 orthologs with respect to translation, viability at host temperatures and pathogenesis. Our results indicate that although the ribosome-binding residues are not fully conserved in LACK, both LACK and TbRACK1 co-sedimented with monosomes and polysomes in LACK/LACK and LACK/TbRACK1 L. major, respectively. LACK/LACK and LACK/TbRACK1 strains differed in their sensitivity to translation inhibitors implying that minor sequence differences between the RACK1 proteins can alter their functional properties. While biochemically distinguishable, both LACK/LACK and LACK/TbRACK1 lines were more tolerant of elevated temperatures, resistant to translation inhibitors, and displayed robust pathogenesis in vivo, contrasting to LACK/- parasites

Disruption of the Putative Ribosome-Binding Motif of a Scaffold Protein Impairs Cytochrome c Oxidase Subunit Expression in Leishmania major

Leishmania parasites are trypanosomatid protozoans that persist in infected human hosts to cause a spectrum of pathologies, from cutaneous and mucocutaneous manifestations to visceral leishmaniasis caused by Leishmania donovani. The latter is usually fatal if not treated. Persistence of L. major in the mammalian host depends upon maintaining gene-regulatory programs to support essential parasite metabolic functions. These include expression and assembly of mitochondrial L. major cytochrome c oxidase (LmCOX) subunits, important for Leishmania ATP production. Significantly, under mammalian conditions, WT levels of LmCOX subunits require threshold levels of the Leishmania ribosome-associated scaffold protein, LACK. Unexpectedly, we find that although disruption of LACK’s putative ribosome-binding motif does not grossly perturb ribosome association or global protein synthesis, it nonetheless impairs COX subunit expression, mitochondrial function, and virulence. Our data indicate that the quality of LACK’s interaction with Leishmania ribosomes is critical for LmCOX subunit expression and parasite mitochondrial function in the mammalian host. Collectively, these findings validate LACK’s ribosomal interactions as a potential therapeutic target.During their parasitic life cycle, through sandflies and vertebrate hosts, Leishmania parasites confront strikingly different environments, including abrupt changes in pH and temperature, to which they must rapidly adapt. These adaptations include alterations in Leishmania gene expression, metabolism, and morphology, allowing them to thrive as promastigotes in the sandfly and as intracellular amastigotes in the vertebrate host. A critical aspect of Leishmania metabolic adaptation to these changes is maintenance of efficient mitochondrial function in the hostile vertebrate environment. Such functions, including generation of ATP, depend upon the expression of many mitochondrial proteins, including subunits of cytochrome c oxidase (COX). Significantly, under mammalian temperature conditions, expression of Leishmania major COX subunit IV (LmCOX4) and virulence are dependent upon two copies of LACK, a gene that encodes the ribosome-associated scaffold protein, LACK (Leishmania ortholog of RACK1 [receptor for activated C kinase]). Targeted replacement of an endogenous LACK copy with a putative ribosome-binding motif-disrupted variant (LACKR34D35G36→LACKD34D35E36) resulted in thermosensitive parasites that showed diminished LmCOX4 expression, mitochondrial fitness, and replication in macrophages. Surprisingly, despite these phenotypes, LACKD34D35E36 associated with monosomes and polysomes and showed no major impairment of global protein synthesis. Collectively, these data suggest that wild-type (WT) LACK orchestrates robust LmCOX4 expression and mitochondrial fitness to ensure parasite virulence, via optimized functional interactions with the ribosome

Early Li-8+beta-NMR investigations in GaAs and Ge

In this paper, we describe initial studies of the structure and dynamics associated with Li-8(+) in bulk crystalline GaAs and Ge. At low temperatures in GaAs, the amplitude of the Li-8(+) resonance signal at approximate to 3 T indicates that a large fraction (at least 70%) of the Li end up in locations with cubic symmetry (i.e. the tetrahedral interstitial and substitutional sites). The linewidth of the beta-NMR Li-8(+) resonance increases dramatically above 150 K, reaches a maximum at about 290 K, and decreases again. This suggests that the Li starts to change its location, probably from an interstitial to a substitutional site, at approximate to 150 K. Experiments in Ge are also described. In this sample, a narrow resonance is seen at low temperatures that is likely due to Li located at an interstitial site. Near room temperature, it appears that Li is converting to another site. (c) 2005 Elsevier B.V. All rights reserved

Depth-controlled beta-NMR of 8Li in a thin silver film.

Depth-controlled beta-NMR can be used to probe the magnetic properties of thin films and interfaces on a nanometer length scale. A 30 keV beam of highly spin-polarized 8Li+ ions was slowed down and implanted into a 50 nm film of Ag deposited on a SrTiO3 substrate. A novel high field beta-NMR spectrometer was used to observe two well resolved resonances which are attributed to Li occupying substitutional and octahedral interstitial sites in the Ag lattice. The temperature dependence of the Knight shifts and spin relaxation rates are consistent with the Korringa law for a simple metal, implying that the NMR of implanted 8Li reflects the spin suspectibility of bulk metallic silver

Depth-controlled beta-NMR of 8Li in a thin silver film.

Depth-controlled beta-NMR can be used to probe the magnetic properties of thin films and interfaces on a nanometer length scale. A 30 keV beam of highly spin-polarized 8Li+ ions was slowed down and implanted into a 50 nm film of Ag deposited on a SrTiO3 substrate. A novel high field beta-NMR spectrometer was used to observe two well resolved resonances which are attributed to Li occupying substitutional and octahedral interstitial sites in the Ag lattice. The temperature dependence of the Knight shifts and spin relaxation rates are consistent with the Korringa law for a simple metal, implying that the NMR of implanted 8Li reflects the spin suspectibility of bulk metallic silver

beta-detected NMR of Li-8 in the normal state of 2H-NbSe2

beta-NMR of isolated Li-8 has been investigated in the normal state of 2H-NbSe2. In a high magnetic field of 3 T a single resonance is observed with a Gaussian line width of 3.5 kHz. The line shape varies weakly as function of magnetic field and temperature but has a strong orientation dependence. The nuclear electric quadrupole splitting is unresolved implying that the electric field gradients are 10-100 times smaller than in other non-cubic crystals. The nuclear spin relaxation rate is also anomalously small but varies linearly with temperature as expected for Korringa relaxation in a metal. These results suggest that Li adopts an interstitial position between the weakly coupled NbSe2 layers and away from the conduction band. (c) 2005 Elsevier B.V. All rights reserved

The new beta-NMR facility at TRIUMF and applications in semiconductors

A new facililty for conducting beta-detected nuclear magnetic resonance (beta-NMR) investigations of condensed matter systems has recently been constucted at TriUniversity Meson Facility in Vancouver, Canada. The unique features of this new facility are described, and some preliminary results on Li-8(+) in GaAs are presented. (C) 2003 Elsevier B.V. All rights reserved