Different domains cooperate to target the human ribosomal L7a protein to the nucleus and to the nucleoli.

The human ribosomal protein L7a is a component of the major ribosomal subunit. We transiently expressed in HeLa cells L7a-β-galactosidase fusion proteins and studied their subcellular localization by indirect immunofluorescence staining with anti-β-galactosidase antibodies. We have identified three distinct domains responsible for the nuclear targeting of the protein: domain I, amino acids 23-51; domain II, amino acids 52-100; domain III, amino acids 101-220, each of which contains at least one nuclear localization signal (NLS). Through subcellular localization analysis of deletion mutants of L7a-β-galactosidase chimeras, we demonstrate that domain II plays a special role because it is necessary, although not sufficient, to target the chimeric β-galactosidase to the nucleoli. In fact, we demonstrate that the nucleolar targeting process requires the presence of domain II plus an additional basic domain that can be represented by an NLS or a basic stretch of amino acids without NLS activity. Thus, when multiple NLS are present, each NLS exerts distinct functions. Domain II drives nucleolar accumulation of a reporter protein with the cooperative action of a short basic amino acid sequence, suggesting a mechanism requiring protein-protein or protein-nucleic acid interactions

Possible, alternative explanations of the T2K observation of the nu_e appearance from an initial nu_mu

An alternative explanation to the emergence of sin^2(2 theta_13) > 0 is discussed. It is pointed out that the recorded T2K events might have been due to some other new physics in the neutrino sector, related to the LSND/MiniBooNE sterile neutrino anomalies, for which there is nowadays a growing evidence. The presently running ICARUS detector with the CNGS beam will be able to distinguish between these two possible sources of the effectComment: 5 pages, 1 figur

cis-acting sequences and trans-acting factors in the localization of mRNA for mitochondrial ribosomal proteins

mRNA localization is a conserved post-transcriptional process crucial for a variety of systems. Although several mechanisms have been identified, emerging evidence suggests that most transcripts reach the protein functional site by moving along cytoskeleton elements. We demonstrated previously that mRNA for mitochondrial ribosomal proteins are asymmetrically distributed in the cytoplasm, and that localization in the proximity of mitochondria is mediated by the 3′-UTR. Here we show by biochemical analysis that these mRNA transcripts are associated with the cytoskeleton through the microtubule network. Cytoskeleton association is functional for their intracellular localization near the mitochondrion, and the 3′-UTR is involved in this cytoskeleton-dependent localization. To identify the minimal elements required for localization, we generated DNA constructs containing, downstream from the GFP gene, deletion mutants of mitochondrial ribosomal protein S12 3′-UTR, and expressed them in HeLa cells. RT-PCR analysis showed that the localization signals responsible for mRNA localization are located in the first 154 nucleotides. RNA pulldown assays, mass spectrometry, and RNP immunoprecipitation assay experiments, demonstrated that mitochondrial ribosomal protein S12 3′-UTR interacts specifically with TRAP1 (tumor necrosis factor receptor-associated protein1), hnRNPM4 (heterogeneous nuclear ribonucleoprotein M4), Hsp70 and Hsp60 (heat shock proteins 70 and 60), and α-tubulin in vitro and in vivo

Constant-q data representation in Neutron Compton scattering on the VESUVIO spectrometer

Standard data analysis on the VESUVIO spectrometer at ISIS is carried out within the Impulse Approximation framework, making use of the West scaling variable y. The experiments are performed using the time-of-flight technique with the detectors positioned at constant scattering angles. Line shape analysis is routinely performed in the y-scaling framework, using two different (and equivalent) approaches: (I) fitting the parameters of the recoil peaks directly to fixed-angle time-of-flight spectral (2) transforming the time-of-flight spectra into fixed-angle y spectra, referred to as the Neutron Compton Profiles, and then fitting the line shape parameters. The present work shows that scattering signals from different fixed-angle detectors can be collected and rebinned to obtain Neutron Compton Profiles at constant wave vector transfer, q, allowing for a suitable interpretation of data in terms of the dynamical structure factor, S(q, w). The current limits of applicability of such a procedure are discussed in terms of the available q-range and relative uncertainties for the VESUVIO experimental set up and of the main approximations involved. (C) 2008 Elsevier B.V. All rights reserved

A new search for anomalous neutrino oscillations at the CERN-PS

The LSND experiment has observed a 3.8 sigma excess of anti-nu_e events from an anti-nu_mu beam coming from pions at rest. If confirmed, the LSND anomaly would imply new physics beyond the standard model, presumably in the form of some additional sterile neutrinos. The MiniBooNE experiment at FNAL-Booster has further searched for the LSND anomaly. Above 475 MeV, the nu_e result is excluding the LSND anomaly to about 1.6 sigma but it introduces an unexplained, new 3.0 sigma anomaly at lower energies, down to 200 MeV. The nu_e data have so far an insufficient statistics to be conclusive with LSND's anti-nu_e. The present proposal at the CERN-PS is based on two strictly identical LAr-TPC detectors in the near and far positions, respectively at 127 and 850 m from the neutrino (or antineutrino) target and focussing horn, observing the electron-neutrino signal. This project will benefit from the already developed technology of ICARUS T600, well tested on surface in Pavia, without the need of any major R&D activity and without the added problems of an underground experiment (CNGS-2). The superior quality of the Liquid Argon imaging TPC and its unique electron - pi-zero discrimination allow full rejection of the NC background, without efficiency loss for electron neutrino detection. In two years of exposure, the far detector mass of 600 tons and a reasonable utilization of the CERN-PS with the refurbished previous TT7 beam line will allow to collect about 10^6 charged current events, largely adequate to settle definitely the LSND anomaly.Comment: 23 pages, 17 figures, added watermark, better referencin

The 3'-untranslated region directs ribosomal protein-encoding mRNAs to specific cytoplasmic regions

mRNA localization is a conserved post-transcriptional process crucial for a variety of systems. We have analyzed the subcellular distribution of mRNAs encoding human cytosolic and mitochondrial ribosomal proteins. Biochemical fractionation experiments showed that the transcripts for cytosolic ribosomal proteins associate preferentially with the cytoskeleton via actin microfilaments. Transfection in HeLa cells of a GFP reporter construct containing the cytosolic ribosomal protein L4 3'-UTR showed that the 3'-UTR is necessary for the association of the transcript to the cytoskeleton. Using confocal analysis we demonstrate that the chimeric transcript is specifically associated with the perinuclear cytoskeleton. We also show that mRNA for mitochondrial ribosomal protein S12 is asymmetrically distributed in the cytoplasm. In fact, this transcript was localized mainly in the proximity of mitochondria, and the localization was 3'-UTR-dependent. In summary, ribosomal protein mRNAs constitute a new class of localized transcripts that share a common localization mechanism

Islet autoimmunity identifies a unique pattern of impaired pancreatic beta-cell function, markedly reduced pancreatic beta cell mass and insulin resistance in clinically diagnosed type 2 diabetes

There is a paucity of literature describing metabolic and histological data in adult-onset autoimmune diabetes. This subgroup of diabetes mellitus affects at least 5% of clinically diagnosed type 2 diabetic patients (T2DM) and it is termed Latent Autoimmune Diabetes in Adults (LADA). We evaluated indexes of insulin secretion, metabolic assessment, and pancreatic pathology in clinically diagnosed T2DM patients with and without the presence of humoral islet autoimmunity (Ab). A total of 18 patients with at least 5-year duration of clinically diagnosed T2DM were evaluated in this study. In those subjects we assessed acute insulin responses to arginine, a glucose clamp study, whole-body fat mass and fat-free mass. We have also analyzed the pancreatic pathology of 15 T2DM and 43 control cadaveric donors, using pancreatic tissue obtained from all the T2DM organ donors available from the nPOD network through December 31, 2013. The presence of islet Ab correlated with severely impaired β-cell function as demonstrated by remarkably low acute insulin response to arginine (AIR) when compared to that of the Ab negative group. Glucose clamp studies indicated that both Ab positive and Ab negative patients exhibited peripheral insulin resistance in a similar fashion. Pathology data from T2DM donors with Ab or the autoimmune diabetes associated DR3/DR4 allelic class II combination showed reduction in beta cell mass as well as presence of autoimmune-associated pattern A pathology in subjects with either islet autoantibodies or the DR3/DR4 genotype. In conclusion, we provide compelling evidence indicating that islet Ab positive long-term T2DM patients exhibit profound impairment of insulin secretion as well as reduced beta cell mass seemingly determined by an immune-mediated injury of pancreatic β-cells. Deciphering the mechanisms underlying beta cell destruction in this subset of diabetic patients may lead to the development of novel immunologic therapies aimed at halting the disease progression in its early stage

Facility for fast neutron irradiation tests of electronics at the ISIS spallation neutron source

The VESUVIO beam line at the ISIS spallation neutron source was set up for neutron irradiation tests in the neutron energy range above 10 MeV. The neutron flux and energy spectrum were shown, in benchmark activation measurements, to provide a neutron spectrum similar to the ambient one at sea level, but with an enhancement in intensity of a factor of 107. Such conditions are suitable for accelerated testing of electronic components, as was demonstrated here by measurements of soft error rates in recent technology field programable gate arrays

The Successful Operation of Hole-type Gaseous Detectors at Cryogenic Temperatures

We have demonstrated that hole-type gaseous detectors, GEMs and capillary plates, can operate up to 77 K. For example, a single capillary plate can operate at gains of above 10E3 in the entire temperature interval between 300 until 77 K. The same capillary plate combined with CsI photocathodes could operate perfectly well at gains (depending on gas mixtures) of 100-1000. Obtained results may open new fields of applications for capillary plates as detectors of UV light and charge particles at cryogenic temperatures: noble liquid TPCs, WIMP detectors or LXe scintillating calorimeters and cryogenic PETs.Comment: Presented at the IEEE Nuclear Science Symposium, Roma, 200