U2 Particles Research Articles

Correlation of Gravitational Waves–Neutrino Detectors: The Long Neutrino Voyage from SN1987A

The correlation between the data obtained with two gravitational wave (GW) detectors (in Rome and in Maryland) and two neutrino detectors (LSD in Italy and Kamiokande in Japan) has been studied in more detail. If we indicate with U1 the response of the gravitational wave detectors and with U2 the response of the neutrino detectors, we find that there is a clear time correlation between U1 and U2: namely U2 comes later than U1 by an amount depending on the energy measured in the neutrino detectors (the delay decreases with the detected neutrino energy). A straightforward calculation would yield a mass for the U2 particles between 4 and 6 eV. If one further assumes that the U2 is due to neutrinos whose mass has been evaluated to be less than 0.1 eV (at 90% c.l.), we hazard the idea that a slow down during the neutrinos’ 168 000 light-year travel from SN1987A to Earth could be inferred.

Characterization of U4 and U6 interactions with the 5' splice site using a S. cerevisiae in vitro trans-splicing system.

Spliceosome assembly has been characterized as the ordered association of the snRNP particles U1, U2, and U4/U6.U5 onto pre-mRNA. We have used an in vitro trans-splicing/cross-linking system in Saccharomyces cerevisiae nuclear extracts to examine the first step of this process, 5' splice site recognition. This trans-splicing reaction has ATP, Mg(2+), and splice-site sequence requirements similar to those of cis-splicing reactions. Using this system, we identified and characterized a novel U4-5' splice site interaction that is ATP-dependent, but does not require the branch point, the 3' splice site, or the 5' end of the U1 snRNA. Additionally, we identified several ATP-dependent U6 cross-links at the 5' splice site, indicating that different regions of U6 sample it before a U6-5' splice site interaction is stabilized that persists through the first step of splicing. This work provides evidence for ATP-dependent U4/U6 association with the 5' splice site independent of ATP-mediated U2 association with the branch point. Furthermore, it defines specific nucleotides in U4 and U6 that interact with the 5' splice site at this early stage, even in the absence of base-pairing with the U1 snRNA.

Characterisation of human and murine snRNP proteins by two-dimensional gel electrophoresis and phosphopeptide analysis of U1-specific 70K protein variants

The proteins of the major human snRNPs U1, U2, U4/U6 and U5 were characterised by two-dimensional electrophoresis, with isoelectric focussing in the first dimension and SDS-polyacrylamide gel electrophoresis in the second. With the exception of protein F, which exhibits an acidic pl value (pl = 3.3), the snRNP proteins are basic. Post-translational modification was found among the proteins associated specifically with the U1 and U2 particles. The most complex modification pattern was observed for the U1-specific 70K protein. This was found in at least 13 isoelectric variants, with pl values ranging from 6.7 to 8.7; these variants differed also in molecular weight. All of the 70K variants are phosphorylated in the cell. Thin-layer analysis of their tryptic phosphopeptides revealed that the 70K variants have four major phosphopeptides in common, in addition to which at least four additional serine residues are phosphorylated to different extents. The comparative phosphopeptide analysis shows that differential phosphorylation alone is not sufficient to explain the occurrence of the many isoelectric variants of 70K, so that the final charge of the 70K variants is determined both by phosphorylation and by other, as yet unidentified posttranslational modifications. By two-dimensional separation of snRNP proteins obtained from mouse Ehrlich ascites tumour cells, it was shown that the pattern of pl values of the mouse proteins was almost identical with the corresponding pattern for human proteins. Even the complex modification patterns of the 70K protein are identical in mouse and man, indicating that the presence in the cell of so many variants of this protein may have functional importance. The major difference between murine and human snRNP proteins is the absence of protein B' from mouse snRNPs. This suggests that the homologous protein B may be able to carry out the task of protein B'.

Mapping of B cell epitopes on small nuclear ribonucleoproteins that react with human autoantibodies as well as with experimentally-induced mouse monoclonal antibodies.

Small nuclear ribonucleoprotein (snRNP) particles are a class of RNA-containing particles in the nucleus of eukaryotic cells. They consist of uridylate-rich small nuclear RNA complexed with several proteins. snRNP particles U1, U2, U4/U6, and U5 all contain a common protein core consisting of proteins B'/B, D, D', E, F, and G. In addition to this core, U1 snRNP particles contain proteins 70K, A, and C, whereas U2 snRNP particles contain proteins A' and B". Almost any of the small nuclear RNA-associated polypeptides is targeted by autoantibodies in the sera from patients with SLE or related connective tissue diseases. We immunized a genetically non-autoimmune mouse with recombinant human B" protein and obtained three mAb reactive with native U2 snRNP particles. Two of these mAb particles cross-reacted with U1 snRNP, 9A9 and 11A1, via epitopes present on the U2 snRNP B" protein as well as on the U1 snRNP-specific A protein. A third mAb 4g3, reacted exclusively with U2 snRNP via a unique epitope on protein B". Two epitopes mapped at the carboxy-terminal region of the B" protein, whereas binding of the third mAb involved both amino- and carboxy-terminal amino acids of the B" protein. Epitope mapping, employing a DNAse I fragment library of the B" cDNA, revealed that the three mAb-reactive sites were discontinuous. Autoantibodies in sera from patients with SLE and other connective tissue diseases competed for binding with the mAb, implying that the mAb define a major autoantibody-reactive region on protein B".

Electron microscopy of U1 small nuclear ribonucleoprotein particles: shape of the particle and position of the 5′ RNA terminus.

Small nuclear ribonucleoprotein particles U1 (U1 snRNPs) from HeLa cells were purified to homogeneity and examined in the electron microscope by negative staining with the double carbon film technique. A survey of several hundred views of the U1 snRNP particles revealed a main body about 8 nm in diameter with two characteristic protuberances 4-7 nm long and 3-4 nm wide. Anti-m3G antibodies were then used to define the site of the m3G base in the 5' cap of the U1 snRNA, close to the site where base-pairing occurs with the 5' splice sequence of pre-mRNA. The m3G base is located on the body of the U1 snRNP, close to one of the protuberances and approximately opposite the other.

Pre-mRNA splicing by complementation with purified human UI, U2, U4/U6 and U5 snRNPs

The four major nucleoplasmic small nuclear ribonucleoprotein particles U1, U2, U4/U6 and U5 can be extensively purified from HeLa cells by immunoaffinity chromatography using a monoclonal anti-trimethylguanosine antibody. The snRNP particles in active splicing extracts are selectively bound to the immunoaffinity matrix, and are then gently eluted by competition with an excess of free nucleoside. Biochemical complementation studies show that the purified snRNPs are active in pre-mRNA splicing, but only in the presence of additional non-snRNP protein factors. All the RNPs that are necessary for splicing can be purified in this manner. The active snRNPs are characterized with respect to their polypeptide composition, and shown to be distinct from several other activities implicated in splicing.

Separation of multiple components of HeLa cell nuclear extracts required for pre-messenger RNA splicing.

Components essential for nuclear pre-messenger RNA splicing have been partially purified from HeLa cell nuclear extracts by chromatography on DEAE-Sepharose, heparin-Sepharose, Mono Q, and Mono S. We have obtained six fractions which, when combined, efficiently splice a synthetic adenovirus 2 major late RNA substrate in vitro. All fractions contain components that support the formation of splicing intermediates (the cleaved 5' exon and the intron-exon 2 lariat). At least one of the fractions also contains an activity that is essential for the second step in the splicing reaction, namely cleavage at the 3' splice site and exon ligation. Two of the fractions are enriched in the major small nuclear ribonucleoprotein particles U1, U2, U4/U6, and U5. They participate in the formation of the splicing complexes which precedes the cleavage and ligation reactions. The remaining four fractions appear to contain protein factors, as suggested by their resistance to micrococcal nuclease.

Fractionation of HeLa cell nuclear extracts reveals minor small nuclear ribonucleoprotein particles.

Upon chromatographic fractionation of HeLa cell nuclear extracts, small RNAs of 145 and 66/65 nucleotides, respectively, were detected that are distinct from the abundant small RNAs present in the extract. These RNAs are precipitated by antibodies directed against the trimethylguanosine cap structure, characteristic for small nuclear RNAs (snRNAs) of the U type. The RNAs of 145 and 66/65 nucleotides appear to be associated with at least one of the proteins common to the major small nuclear ribonucleoprotein particles U1 to U6, since they are specifically bound by anti-Sm antibodies. These criteria characterize the RNAs that are 145 and 66/65 nucleotides in length as U-type snRNAs. Upon gel filtration, the RNAs are found within particles of molecular weights approximately equal to 150,000 and 115,000, respectively. The RNA of 145 nucleotides represents a different minor snRNA, designated U11, whereas the RNA of 66/65 nucleotides may correspond to either mammalian U7 or U10 RNA.

Fragmentation of Uranium Dioxide after Molten Uranium Dioxide-Sodium Interaction

A series of 30 runs were performed to gain information on the fragmentation of molten UO2when dropped into a bath of liquid sodium. In this experiment, hollow cylindrical UO2 pellets were heated with electric current fed to a tungsten rod, passed through to center hole. The resulting molten UO2 dripped down its fall into the sodium bath. High-speed cinematography (500~2,000 frames/sec) was utilized to observe the fragmentation of the UO2 droplets upon its impact with the sodium surface. Records were taken of the changes in pressure and temperature induced in the sodium bath by the impact of the U2 droplets. The physical characteristics of the solid U2 particles resulting from the quench were examined for particle size distribution, particle shape and surface condition, as well as microstructure, to gain an insight into the mechanism of fragmentation. It is concluded from the study that, for the purpose of Fuel Coolant Interaction (FCI) analysis, the U2 particles can be assumed to be spherical in shape and...

Fragmentation of Uranium Dioxide after Molten Uranium Dioxide-Sodium Interaction

A series of 30 runs were performed to gain information on the fragmentation of molten UO2when dropped into a bath of liquid sodium. In this experiment, hollow cylindrical UO2 pellets were heated with electric current fed to a tungsten rod, passed through to center hole. The resulting molten UO2 dripped down its fall into the sodium bath. High-speed cinematography (500~2,000 frames/sec) was utilized to observe the fragmentation of the UO2 droplets upon its impact with the sodium surface. Records were taken of the changes in pressure and temperature induced in the sodium bath by the impact of the U2 droplets. The physical characteristics of the solid U2 particles resulting from the quench were examined for particle size distribution, particle shape and surface condition, as well as microstructure, to gain an insight into the mechanism of fragmentation. It is concluded from the study that, for the purpose of Fuel Coolant Interaction (FCI) analysis, the U2 particles can be assumed to be spherical in shape and to have log-normal particle-size distribution. The density function in terms of particle weight, representing the finest particle-size distribution, can be expressed on the basis of presently available empirical data, by , D: Diameter of U2 particle in microns.