Serine/arginine-rich splicing factor 3 (SRSF3) likely has wide-ranging roles in gene expression and facilitation of tumor cell growth. SRSF3 knockdown induced G1 arrest and apoptosis in colon cancer ...cells (HCT116) in association with altered expression of 833 genes. Pathway analysis revealed 'G1/S Checkpoint Regulation' as the most highly enriched category in the affected genes. SRSF3 knockdown did not induce p53 or stimulate phosphorylation of p53 or histone H2A.X in wild-type HCT116 cells. Furthermore, the knockdown induced G1 arrest in p53-null HCT116 cells, suggesting that p53-dependent DNA damage responses did not mediate the G1 arrest. Real-time reverse transcription-polymerase chain reaction and western blotting confirmed that SRSF3 knockdown reduced mRNA and protein levels of cyclins (D1, D3 and E1), E2F1 and E2F7. The decreased expression of cyclin D and E2F1 likely impaired the G1-to-S-phase progression. Consequently, retinoblastoma protein remained hypophosphorylated in SRSF3 knockdown cells. The knockdown also induced apoptosis in association with reduction of BCL2 protein levels. We also found that SRSF3 knockdown facilitated skipping of 81 5'-nucleotides (27 amino acids) from exon 8 of homeodomain-interacting protein kinase-2 (HIPK2) and produced a HIPK2 Δe8 isoform. Full-length HIPK2 (HIPK2 FL) is constantly degraded through association with an E3 ubiquitin ligase (Siah-1), whereas HIPK2 Δe8, lacking the 27 amino acids, lost Siah-1-binding ability and became resistant to proteasome digestion. Interestingly, selective knockdown of HIPK2 FL induced apoptosis in various colon cancer cells expressing wild-type or mutated p53. Thus, these findings disclose an important role of SRSF3 in the regulation of the G1-to-S-phase progression and alternative splicing of HIPK2 in tumor growth.
RNA-binding proteins and microRNAs are potent post-transcriptional regulators of gene expression. Human transformer 2β (Tra2β) is a serine/arginine-rich-like protein splicing factor and is now ...implicated to have wide-ranging roles in gene expression as an RNA-binding protein. RNA immunoprecipitation (RIP) with an anti-Tra2β antibody and microarray analysis identified a subset of Tra2β-associated mRNAs in HCT116 human colon cancer cells, many of which encoded cell death-related proteins including Bcl-2 (B-cell CLL/lymphoma 2). Tra2β knockdown in HCT116 cells decreased Bcl-2 expression and induced apoptosis. Tra2β knockdown accelerated the decay of BCL2α mRNA that encodes Bcl-2 and full-length 3' UTR, while it did not affect the stability of BCL2β mRNA having a short, alternatively spliced 3' UTR different from BCL2α 3' UTR. RIP assays with anti-Tra2β and anti-Argonaute 2 antibodies, respectively, showed that Tra2β bound to BCL2α 3' UTR, and that Tra2β knockdown facilitated association of miR-204 with BCL2α 3' UTR. The consensus sequence (GAA) for Tra2β-binding lies within the miR-204-binding site of BCL2 3' UTR. Mutation of the consensus sequence canceled the binding of Tra2β to BCL2 3' UTR without disrupting miR-204-binding to BCL2 3' UTR. Transfection of an anti-miR-204 or introduction of three-point mutations into the miR-204-binding site increased BCL2 mRNA and Bcl-2 protein levels. Inversely, transfection of precursor miR-204 reduced their levels. Experiments with Tra2β-silenced or overexpressed cells revealed that Tra2β antagonized the effects of miR-204 and upregulated Bcl-2 expression. Furthermore, TRA2β mRNA expression was significantly upregulated in 22 colon cancer tissues compared with paired normal tissues and positively correlated with BCL2 mRNA expression. Tra2β knockdown in human lung adenocarcinoma cells (A549) increased their sensitivity to anticancer drugs. Taken together, our findings suggest that Tra2β regulates apoptosis by modulating Bcl-2 expression through its competition with miR-204. This novel function may have a crucial role in tumor growth.
In August 2015, the CALorimetric Electron Telescope (CALET), designed for long exposure observations of high energy cosmic rays, docked with the International Space Station (ISS) and shortly ...thereafter began to collect data. CALET will measure the cosmic ray electron spectrum over the energy range of 1 GeV to 20 TeV with a very high resolution of 2% above 100 GeV, based on a dedicated instrument incorporating an exceptionally thick 30 radiation-length calorimeter with both total absorption and imaging (TASC and IMC) units. Each TASC readout channel must be carefully calibrated over the extremely wide dynamic range of CALET that spans six orders of magnitude in order to obtain a degree of calibration accuracy matching the resolution of energy measurements. These calibrations consist of calculating the conversion factors between ADC units and energy deposits, ensuring linearity over each gain range, and providing a seamless transition between neighboring gain ranges. This paper describes these calibration methods in detail, along with the resulting data and associated accuracies. The results presented in this paper show that a sufficient accuracy was achieved for the calibrations of each channel in order to obtain a suitable resolution over the entire dynamic range of the electron spectrum measurement.
The CALorimetric Electron Telescope (CALET), launched for installation on the International Space Station (ISS) in August, 2015, has been accumulating scientific data since October, 2015. CALET is ...intended to perform long-duration observations of high-energy cosmic rays onboard the ISS. CALET directly measures the cosmic-ray electron spectrum in the energy range of 1 GeV to 20 TeV with a 2% energy resolution above 30 GeV. In addition, the instrument can measure the spectrum of gamma rays well into the TeV range, and the spectra of protons and nuclei up to a PeV.
In order to operate the CALET onboard ISS, JAXA Ground Support Equipment (JAXA-GSE) and the Waseda CALET Operations Center (WCOC) have been established at JAXA and Waseda University, respectively. Scientific operations using CALET are planned at WCOC, taking into account orbital variations of geomagnetic rigidity cutoff. Scheduled command sequences are used to control the CALET observation modes on orbit. Calibration data acquisition by, for example, recording pedestal and penetrating particle events, a low-energy electron trigger mode operating at high geomagnetic latitude, a low-energy gamma-ray trigger mode operating at low geomagnetic latitude, and an ultra heavy trigger mode, are scheduled around the ISS orbit while maintaining maximum exposure to high-energy electrons and other high-energy shower events by always having the high-energy trigger mode active. The WCOC also prepares and distributes CALET flight data to collaborators in Italy and the United States.
As of August 31, 2017, the total observation time is 689 days with a live time fraction of the total time of ∼ 84%. Nearly 450 million events are collected with a high-energy (E > 10 GeV) trigger. In addition, calibration data acquisition and low-energy trigger modes, as well as an ultra-heavy trigger mode, are consistently scheduled around the ISS orbit. By combining all operation modes with the excellent-quality on-orbit data collected thus far, it is expected that a five-year observation period will provide a wealth of new and interesting results.
The CALorimetric Electron Telescope (CALET)on the International Space Station consists of a high-energy cosmic-ray CALorimeter (CAL)and a lower-energy CALET Gamma-ray Burst Monitor (CGBM). CAL is ...sensitive to electrons up to 20 TeV, cosmic-ray nuclei from Z=1 throughZ∼40, and gamma rays over the range1 GeV–10 TeV. CGBM observes gamma rays from 7 keV to 20 MeV. The combined CAL-CGBM instrument has conducted a search for gamma-ray bursts (GRBs)since 2015 October. We report here on the results of a search for X-ray/gamma-ray counterparts to gravitational-wave events reported during the LIGO/Virgo observing run O3. No events have been detected that pass all acceptance criteria. We describe the components, performance, and triggering algorithms of the CGBM—the two Hard X-ray Monitors consisting of LaBr3(Ce)scintillators sensitive to 7 keV–1 MeV gamma rays and a Soft Gamma-ray Monitor BGO scintillator sensitive to 40 keV–20 MeV—and the high-energy CAL consisting of a charge detection module, imaging calorimeter, and the fully active total absorption calorimeter. The analysis procedure is described and upper limits to the time-averaged fluxes are presented.
Homeodomain-interacting protein kinase 2 (HIPK2) is a potential tumor suppressor that has a crucial role in the DNA damage response (DDR) by regulating cell-cycle checkpoint activation and apoptosis. ...However, it is unclear whether HIPK2 exerts distinct roles in DNA damage repair. The aim of this study was to identify novel target molecule(s) of HIPK2, which mediates HIPK2-dependent DNA damage repair. HIPK2-knockdown human colon cancer cells (HCT116) or hipk1/hipk2 double-deficient mouse embryonic fibroblasts could not remove histone H2A.X phosphorylated at Ser139 (γH2A.X) after irradiation with a sublethal dose (10 J/m(2)) of ultraviolet (UV)-C, resulting in apoptosis. Knockdown of HIPK2 in p53-null HCT116 cells similarly promoted the UV-C-induced γH2A.X accumulation and apoptosis. Proteomic analysis of HIPK2-associated proteins using liquid chromatography-tandem mass spectrometry identified heterochromatin protein 1γ (HP1γ) as a novel target for HIPK2. Immunoprecipitation experiments with HCT116 cells expressing FLAG-tagged HIPK2 and one of the HA-tagged HP1 family members demonstrated that HIPK2 specifically associated with HP1γ, but not with HP1α or HP1β, through its chromo-shadow domain. Mutation of the HP1box motif (883-PTVSV-887) within HIPK2 abolished the association. HP1γ knockdown also enhanced accumulation of γH2A.X and apoptosis after sublethal UV-C irradiation. In vitro kinase assay demonstrated an HP1γ-phosphorylating activity of HIPK2. Sublethal UV-C irradiation phosphorylated HP1γ. This phosphorylation was absent in endogenous HIPK2-silenced cells with HIPK2 3'UTR siRNA. Overexpression of FLAG-HIPK2, but not the HP1box-mutated or kinase-dead HIPK2 mutant, in the HIPK2-silenced cells increased HP1γ binding to trimethylated (Lys9) histone H3 (H3K9me3), rescued the UV-C-induced phosphorylation of HP1γ, triggered release of HP1γ from histone H3K9me3 and suppressed γH2A.X accumulation. Our results suggest that HIPK2-dependent phosphorylation of HP1γ may participate in the regulation of dynamic interaction between HP1γ and histone H3K9me3 to promote DNA damage repair. This HIPK2/HP1γ pathway may uncover a new functional aspect of HIPK2 as a tumor suppressor.
We report on the measurements performed with relativistic ions from Be to Fe, at the Fragment Separator (FRS) of the GSI Helmholtzzentrum für Schwerionenforschung in Darmstadt, to test the ...performance of charge-sensitive detectors that were designed to separate – via multiple dE/dx measurements – fully stripped nuclei of cosmic origin in the experiment CALET. The latter is a space mission by the Japanese Space Agency (JAXA) scheduled to be launched to the International Space Station (ISS) in 2013. The CALET instrument is managed by an international collaboration and it is scheduled to take data for 5 years on the Exposure Facility (JEM-EF) of the Japanese module KIBO on the ISS.
The aim of the test was to accurately measure the response of the scintillator to different nuclear species and parametrize the saturation of the scintillation light in order to assess the impact of this effect on the charge resolution of the instrument.
► Charge identification of relativistic cosmic nuclei. ► Saturation of scintillation light from ionization by heavy nuclei. ► Charge resolution with scintillators with high Z ionizing radiation.
We present the results of searches for gamma-ray counterparts of the LIGO/Virgo gravitational wave events using CALorimetric Electron Telescope (CALET) observations. The main instrument of CALET, ...CALorimeter (CAL), observes gamma-rays from ∼1 GeV up to 10 TeV with a field of view (FOV) of nearly 2 sr. In addition, the CALET gamma-ray burst monitor views ∼3 sr and ∼2π sr of the sky in the 7 keV-1 MeV and the 40 keV-20 MeV bands, respectively, by using two different crystal scintillators. The CALET observations on the International Space Station started in 2015 October, and here we report analyses of events associated with the following gravitational wave events: GW151226, GW170104, GW170608, GW170814, and GW170817. Although only upper limits on gamma-ray emission are obtained, they correspond to a luminosity of 1049 ∼ 1053 erg s−1 in the GeV energy band depending on the distance and the assumed time duration of each event, which is approximately on the order of luminosity of typical short gamma-ray bursts. This implies that there will be a favorable opportunity to detect high-energy gamma-ray emission in further observations if additional gravitational wave events with favorable geometry will occur within our FOV. We also show the sensitivity of CALET for gamma-ray transient events, which is on the order of 10−7 erg cm−2 s−1 for an observation of 100 s in duration.
The CALorimetric Electron Telescope primary detector (CALET-CAL) is a 30 radiation-length-deep hybrid calorimeter designed for the accurate measurement of high-energy cosmic rays. It is capable of ...triggering on and giving near complete containment of electromagnetic showers from primary electrons and gamma rays from 1 GeV to over 10 TeV. The first 24 months of on-orbit scientific data (2015 November 01-2017 October 31) provide valuable characterization of the performance of the calorimeter based on analyses of the gamma-ray data set in general and bright point sources in particular. We describe the gamma-ray analysis, the expected performance of the calorimeter based on Monte Carlo simulations, the agreement of the flight data with the simulated results, and the outlook for long-term gamma-ray observations with the CAL.
ABSTRACT We present upper limits in the hard X-ray and gamma-ray bands at the time of the Laser Interferometer Gravitational-wave Observatory (LIGO) gravitational-wave event GW151226 derived from the ...CALorimetric Electron Telescope (CALET) observation. The main instrument of CALET, CALorimeter (CAL), observes gamma-rays from ∼1 GeV up to 10 TeV with a field of view of ∼2 sr. The CALET gamma-ray burst monitor (CGBM) views ∼3 sr and ∼2π sr of the sky in the 7 keV-1 MeV and the 40 keV-20 MeV bands, respectively, by using two different scintillator-based instruments. The CGBM covered 32.5% and 49.1% of the GW151226 sky localization probability in the 7 keV-1 MeV and 40 keV-20 MeV bands respectively. We place a 90% upper limit of 2 × 10−7 erg cm−2 s−1 in the 1-100 GeV band where CAL reaches 15% of the integrated LIGO probability (∼1.1 sr). The CGBM 7 upper limits are 1.0 × 10−6 erg cm−2 s−1 (7-500 keV) and 1.8 × 10−6 erg cm−2 s−1 (50-1000 keV) for a 1 s exposure. Those upper limits correspond to the luminosity of 3-5 × 1049 erg s−1, which is significantly lower than typical short GRBs.
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