Temporal regulation of murine cytomegalovirus transcription and mapping of viral RNA synthesized at immediate early times after infection

The replication of murine cytomegalovirus strain Smith in murine embryonic fibroblasts was investigated at immediate early, early, and late times after infection. Cloned subgenomic HindIII fragments of murine cytomegalovirus DNA served to define the regions of transcription. At immediate early times viral RNA classes ranging in size from 5.1 to 1.05 kilobases (kb) were transcribed mainly from the fragments HindIII-K and -L, whereas low levels of transcription were detected from the two termini HindIII-E and HindIII-N. A characteristic pattern of proteins could be translated from immediate early RNA in vitro. At early and late times after infection transcription from all HindIII fragments occurred, but different patterns of transcripts and proteins could be identified. Inhibitors of DNA synthesis induced differences in the late transcription pattern, located in the HindIII-F fragment. The coding region for abundant immediate early transcription could be located at between 0.769 and 0.817 map units. A plasmic clone containing the main part (0.769 to 0.815 map units) of this region was constructed. This region coded for six polyadenylated immediate early RNA species of 5.1, 2.75, 2.0, 1.75, 1.65, and 1.05 kb in size. Only the 1.75-kb RNA originated entirely from the HindIII-L fragment. The 5.1- and 2.75-kb RNA species were encoded by both the HindIII-L and HindIII-K fragments, and the 2.0-, 1.65-, and 1.05-kb RNA species were entirely transcribed within HindIII-K

Nonperturbative analysis of coupled quantum dots in a phonon bath

Transport through coupled quantum dots in a phonon bath is studied using the recently developed real-time renormalization-group method. Thereby, the problem can be treated beyond perturbation theory regarding the complete interaction. A reliable solution for the stationary tunnel current is obtained for the case of moderately strong couplings of the dots to the leads and to the phonon bath. Any other parameter is arbitrary, and the complete electron-phonon interaction is taken into account. Experimental results are quantitatively reproduced by taking into account a finite extension of the wavefunctions within the dots. Its dependence on the energy difference between the dots is derived.Comment: 8 pages, 6 figure

Location, transcripts, and translation products

Cloned genomic fragments from the region (0.769 to 0.818 map units) coding for immediate-early (IE) transcripts of murine cytomegalovirus (MCMV) were used to analyze the physical organization of this region, the direction of transcription, and the proteins synthesized in vitro. Three IE transcription units could be identified. From IE coding region 1 (ie1; 0.781 to 0.796 map units) a dominant 2.75-kilobase (kb) RNA was transcribed from right to left on the prototype arrangement of the MCMV genome which directed the synthesis of an 89,000-molecular-weight polypeptide (89K polypeptide), the major IE protein. This phosphoprotein (pp89) has been shown to be active in the regulation of transcription. Upstream of ie1 and separated by the MCMV enhancer sequence was a second IE coding region, ie2, which was mapped at 0.803 to 0.817 map units. From ie2 a 1.75-kb RNA of moderate abundance was transcribed in the direction opposite to that of the ie1 RNA. After hybrid selection of the ie2 transcript, a 43,000-molecular-weight translation product was detected. A third coding region, ie3, was located directly downstream of ie1 (0.773 to 0.781 map units). The series of RNAs with low abundance, terminating in ie3, probably used the ie1 transcription start site and ranged from 1.0 to 5.1 kb in size. The 5.1-kb RNA apparently represents the nonspliced transcript from both coding regions ie1 and ie3. A 15K polypeptide was translated in vitro from RNA that was hybrid selected by ie3 sequences. Immunoprecipitation with monoclonal antibody revealed that 31K to 67K polypeptides were related to pp89. Some of these proteins were translated from RNAs that were smaller than 2.75 kb. Polypeptides related to pp89 were also synthesized in vivo. Because polypeptides unrelated to pp89 that were translated from RNA that was selected by ie2 and ie3 sequences were not immunoprecipitated by murine antisera, we assumed that the amount of these proteins synthesized in vivo during infection was probably very lo

ATLAS Pixel Detector: Running Experience

The ATLAS Pixel Detector is the innermost tracking device of the ATLAS Experiment. It was successfully commissioned in the year 2008. Calibration measurements have been performed in-situ to set and measure important detector parameters like threshold, charge measurement calibration and timewalk. In combined data taking with the other ATLAS subdetectors by now more than half a million cosmic ray tracks have been collected, which have been used to perform a first alignment and to measure pixel efficiency and noise occupancy. This paper presents results both from the calibration effort in 2008 and from data taking with cosmic rays. A summary of the current detector status is given

Comparison of lunar rocks and meteorites: Implications to histories of the moon and parent meteorite bodies

A number of similarities between lunar and meteoritic rocks are reported and suggest that the comparison is essential for a clear understanding of meteorites as probes of the early history of the solar systems: (1) Monomict and polymict breccias occur in lunar rocks, as well as in achondritic and chondritic meteorites, having resulted from complex and repeated impact processes. (2) Chondrules are present in lunar, as well as in a few achondritic and most chondritic meteorites. It is pointed out that because chondrules may form in several different ways and in different environments, a distinction between the different modes of origin and an estimate of their relative abundance is important if their significance as sources of information on the early history of the solar system is to be clearly understood. (3) Lithic fragments are very useful in attempts to understand the pre- and post-impact history of lunar and meteoritic breccias. They vary from little modified (relative to the apparent original texture), to partly or completely melted and recrystallized lithic fragments

The cytolytic T lymphocyte response to the murine cytomegalovirus

Limiting dilution (LD) analysis with two modifications, the expansion and the restimulation LD assay, led to the detection and quantification of two distinct in vivo maturation stages within the lineage of virus- specific self-restricted CTL after infection of mice with the murine cytomegalovirus (MCMV). A low frequency set, representing an average of 15% of the specifically activated CTL-P in a draining lymph node, generated virus-specific lytic activity in the absence of antigen, solely under expansion conditions provided by growth and differentiation interleukins. These cells were considered to be active and were denoted antigen-independent or interleukin-receptive CTL-P (IL- CTL-P). A high frequency set required additional antigen in vitro to generate functionally active clones, and therefore the cells were termed antigen-dependent. Both sets are present in vivo simultaneously at the peak of the acute immune response and represent antigen- activated cells because their existence strictly depends on a preceding priming event. IL-CTL-P disappear quickly after acute infection and are absent during the memory state. It is proposed that the isolation of IL- CTL-P could serve to detect viral antigen expression during persistent and/or recurrent herpes virus infections

Characterization of the murine cytomegalovirus early transcription unit e1 that is induced by immediate-early proteins

The regulation of murine cytomegalovirus early (E) gene expression was studied in the cell line B25, which is stably transfected with the immediate-early ie1/ie3 gene complex. Infection of B25 cells in the presence of the protein synthesis inhibitor cycloheximide resulted in the expression of some E genes, whereas for the expression of other E genes prior protein synthesis was still mandatory, thus showing differences in the expression requirements of individual E genes. Transcription unit e1, a member of the E genes induced by immediate-early products of the ie1/ie3 gene complex, was characterized. It is located between map units 0.709 and 0.721 of the genome of murine cytomegalovirus strain Smith. A 2.6-kilobase RNA specified in this region is spliced from three exons of 912, 177, and 1,007 or 1,020 nucleotides, which are separated by introns of 93 and 326 nucleotides. The second AUG located in the first exon 119 nucleotides downstream of the 5' cap site is followed by an open reading frame of 990 nucleotides. The predicted polypeptide of 330 amino acids has a calculated molecular mass of 36.4 kilodaltons. Transfection with e1 revealed three antigenically related proteins of 36, 37, and 38 kilodaltons; these proteins probably represent differently modified forms of the predicted protein. These three proteins are phosphorylated and are associated with intranuclear inclusion bodies. A 33-kilodalton protein also derived from e1 was identified as a product of nonspliced transcripts. Comparison of amino acid sequences revealed homology between the murine cytomegalovirus transcription unit e1 and a human cytomegalovirus E transcription unit

Supernova neutrinos: Flavor-dependent fluxes and spectra

Transporting nu_mu and nu_tau in a supernova (SN) core involves several processes that have been neglected in traditional simulations. Based on a Monte Carlo study we find that the flavor-dependent spectral differences are much smaller than is often stated in the literature. A full-scale SN simulation using a Boltzmann solver and including all relevant neutrino reactions confirms these results. The flavor-dependent flux differences are largest during the initial accretion phase.Comment: Proceedings NOON 03, Kanazawa, 10-14 Feb 200

Experiment K-6-12. Morphometric studies of atrial or granules and hepatocytes. Part 1: Morphometric study of the liver; Part 2: The atrial granular accumulations

The livers of flight, F, rats from the Cosmos 1887 mission were markedly paler and heavier than those of the synchronous, S, and vivarium, V, controls. In the F group, microscopic study revealed extensive hepatocytic intracytoplasmic vacuolization which was moderate in the S and minimal in the V groups. The vacuoles were not sudanophilic and therefore were regarded as glycogenic in origin. To obtain objective data concerning the extent of the vacuolization, livers were examined by computer assisted morphometry. Measurements of profile area and perimeter of the hepatocyte nuclei and vacuoles were evaluated according to stereological principles. Results indicated that the volume density of the nuclei was less in the F group than in the S(p equal less than 0.0002) and V(p equal less than 0.001) groups. Mean volume of individual nuclei did not differ. Volume density of the vacuoles was greater in the F than in the V group (p equal less than 0.02) while their mean diameter was less (p equal less than 0.05). To ascertain the relationship between increase in liver weight of the flight animals and the results of this study, an assumption was made that the specific gravity of the vacuolar contents was similar to the other extranuclear components of the hepatocyte. On that basis, calculations showed that the elevated vacuolar volume density in the flight group did not cause the increased liver weight in those animals, but that the non-nuclear, non-vacuolar parenchymal compartment did contribute significantly. Factors that may have played a causal role in liver weight and vacuolar compartment increases are discussed