Trans-activation of the human SOX3 promoter by MAZ in NT2/D1 cells

U ovom radu proučavana je uloga tri visoko konzervisana potencijalna mesta vezivanja za "Myc-associated zinc finger protein" (MAZ) u regulaciji ekspresije humanog SOX3 gena. Eseji izmenjene elektroforetske pokretljivosti u prisustvu antitela na MAZ ukazuju da kompleksi koji se formiraju na dva od tri proučavana mesta u okviru SOX3 promotora sadrže MAZ protein. Takođe, u eksperimentima kotransfekcije smo pokazali da MAZ ima ulogu pozitivnog regulatora transkripcije SOX3 gena, kako u nediferenciranim, tako i u diferenciranim NT2/D1 ćelijama. Iako je MAZ povećao i bazalnu i retinoičnom kiselinom indukovanu promotorsku aktivnost, naši rezultati ukazuju da ovaj transkripcioni faktor ne doprinosi inducibilnosti SOX3 promotora tokom neuralne diferencijacije u prisustvu retinoične kiseline.In this study, we examine the role of three highly conserved putative binding sites for Myc-associated zinc finger protein (MAZ) in regulation of the human SOX3 gene expression. Electrophoretic mobility shift and supershift assays indicate that complexes formed at two out of three MAZ sites of the human SOX3 promoter involve ubiquitously expressed MAZ protein. Furthermore, in cotransfection experiments we demonstrate that MAZ acts as a positive regulator of SOX3 gene transcription in both undifferentiated and RA-differentiated NT2/D1 cells. Although MAZ increased both basal and RA-induced promoter activity, our results suggest that MAZ does not contribute to RA inducibility of the SOX3 promoter during neuronal differentiation of NT2/D1 cells

Suppression of cell cycle progression by Jun dimerization protein (JDP2) involves down-regulation of cyclin A2

We report here a novel role for Jun dimerization protein-2 (JDP2) as a regulator of the progression of normal cells through the cell cycle. To determine the role of JDP2 in vivo, we generated Jdp2 knock-out (Jdp2KO) mice by targeting exon 1 to disrupt the site of initiation of transcription. The healing of wounded skin of Jdp2KO mice proceeded more rapidly than that of control mice and more proliferating cells were found at wound margins. Fibroblasts derived from embryos of Jdp2KO mice proliferated more rapidly and formed more colonies than wild-type fibroblasts. JDP2 was recruited to the promoter of the gene for cyclin A2 (ccna2) at a previously unidentified AP-1 site. Cells lacking Jdp2 had elevated levels of cyclin A2 mRNA. Moreover, reintroduction of JDP2 resulted in repression of transcription of ccna2 and of cell cycle progression. Thus, transcription of the gene for cyclin A2 appears to be a direct target of JDP2 in the suppression of cell proliferation

Characteristics of Anemone Active Regions Appearing in Coronal Holes Observed with {\it Yohkoh} Soft X-ray Telescope

Coronal structure of active regions appearing in coronal holes is studied by using the data obtained with the Soft X-Ray Telescope (SXT) aboard {\it Yohkoh} from 1991 November to 1993 March. The following characteristics are found; Many of active regions appearing in coronal holes show a structure that looks like a ``sea-anemone''. Such active regions are called {\it anemone ARs}. About one-forth of all active regions that were observed with SXT from their births showed the anemone structure. For almost all the anemone ARs, the order of magnetic polarities is consistent with the Hale-Nicholson's polarity law. These anemone ARs also showed more or less east-west asymmetry in X-ray intensity distribution, such that the following (eastern) part of the ARs is brighter than its preceding (western) part. This, as well as the anemone shape itself, is consistent with the magnetic polarity distribution around the anemone ARs. These observations also suggest that an active region appearing in coronal holes has simpler (less sheared) and more preceding-spot-dominant magnetic structure than those appearing in other regions.Comment: 11 pages, 3 tables, 4 figure

Identification of mouse Jun dimerization protein 2 as a novel repressor of ATF-211The nucleotide sequence reported herein has been deposited in the DDBJ, EMBL and GenBank databanks under the accession number AB034697.

AbstractA mouse cDNA that encodes a DNA-binding protein was identified by yeast two-hybrid screening, using activating transcription factor-2 (ATF-2) as the bait. The protein contained a bZIP (basic amino acid-leucine zipper region) domain and its amino acid sequence was almost identical to that of rat Jun dimerization protein 2 (JDP2). Mouse JDP2 interacted with ATF-2 both in vitro and in vivo via its bZIP domain. It was encoded by a single gene and various transcripts were expressed in all tested tissues of adult mice, as well as in embryos, albeit at different levels in various tissues. Furthermore, mouse JDP2 bound to the cAMP-response element (CRE) as a homodimer or as a heterodimer with ATF-2, and repressed CRE-dependent transcription that was mediated by ATF-2. JDP2 was identified as a novel repressor protein that affects ATF-2-mediated transcription

Magnetic Reconnection Triggered by the Parker Instability in the Galaxy: Two-Dimensional Numerical Magnetohydrodynamic Simulations and Application to the Origin of X-Ray Gas in the Galactic Halo

We propose the Galactic flare model for the origin of the X-ray gas in the Galactic halo. For this purpose, we examine the magnetic reconnection triggered by Parker instability (magnetic buoyancy instability), by performing the two-dimensional resistive numerical magnetohydrodynamic simulations. As a result of numerical simulations, the system evolves as following phases: Parker instability occurs in the Galactic disk. In the nonlinear phase of Parker instability, the magnetic loop inflates from the Galactic disk into the Galactic halo, and collides with the anti-parallel magnetic field, so that the current sheets are created in the Galactic halo. The tearing instability occurs, and creates the plasmoids (magnetic islands). Just after the plasmoid ejection, further current-sheet thinning occurs in the sheet, and the anomalous resistivity sets in. Petschek reconnection starts, and heats the gas quickly in the Galactic halo. It also creates the slow and fast shock regions in the Galactic halo. The magnetic field ($B\sim 3 \mu$G), for example, can heat the gas ($n\sim 10^{-3}$ cm$^{-3}$) to temperature of $\sim 10^6$ K via the reconnection in the Galactic halo. The gas is accelerated to Alfv\'en velocity ($\sim 300$ km s$^{-1}$). Such high velocity jets are the evidence of the Galactic flare model we present in this paper, if the Doppler shift of the bipolar jet is detected in the Galactic halo. Full size figures are available at http://www.kwasan.kyoto-u.ac.jp/~tanuma/study/ApJ2002/ApJ2002.htmlComment: 13 pages, 12 figures, uses emulateapj.sty, accepted by Ap

Jun Dimerization Protein 2 Controls Senescence and Differentiation via Regulating Histone Modification

Transcription factor, Jun dimerization protein 2 (JDP2), binds directly to histones and DNAs and then inhibits the p300-mediated acetylation both of core histones and of reconstituted nucleosomes that contain JDP2 recognition DNA sequences. JDP2 plays a key role as a repressor of adipocyte differentiation by regulation of the expression of the gene C/EBPδ via inhibition of histone acetylation. Moreover, JDP2-deficient mouse embryonic fibroblasts (JDP2−/− MEFs) are resistant to replicative senescence. JDP2 inhibits the recruitment of polycomb repressive complexes (PRC1 and PRC2) to the promoter of the gene encoding p16Ink4a, resulting from the inhibition of methylation of lysine 27 of histone H3 (H3K27). Therefore, it seems that chromatin-remodeling factors, including the PRC complex controlled by JDP2, may be important players in the senescence program. The novel mechanisms that underline the action of JDP2 in inducing cellular senescence and suppressing adipocyte differentiation are reviewed

A HR-like Diagram for Solar/Stellar Flares and Corona -- Emission Measure vs Temperature Diagram

In our previous paper, we have presented a theory to explain the observed universal correlation between the emission measure ($EM=n^2 V$) and temperature (T) for solar/stellar flares on the basis of the magnetic reconnection model with heat conduction and chromospheric evaporation. Here n is the electron density and V is the volume. By extending our theory to general situations, we examined the EM-T diagram in detail, and found the following properties: 1) The universal correlation sequence (``main sequence flares'') with $EM \propto T^{17/2}$ corresponds to the case of constant heating flux or equivalently the case of constant magnetic field strength in the reconnection model. 2) The EM-T diagram has a forbidden region, where gas pressure of flares exceeds magnetic pressure. 3) There is a coronal branch with $EM \propto T^{15/2}$ for $T< 10^7$ K, and $EM \propto T^{13/2}$ for $T> 10^7$ K. This branch is situated left side of the main sequence flares in the EM-T diagram. 4) There is another forbidden region determined by the length of flare loop; a lower limit of flare loop is $10^7$ cm. Small flares near this limit correspond to nanoflares observed by SOHO/EIT. 5) We can plot flare evolution track on the EM-T diagram. A flare evolves from the coronal branch to main sequence flares, then returns to the coronal branch eventually. These properties of the EM-T diagram are similar to those of the HR diagram for stars, and thus we propose that the EM-T diagram is quite useful to estimate the physical quantities (loop length, heating flux, magnetic field strength, total energy and so on) of flares and corona when there is no spatially resolved imaging observations.Comment: 31 pages, ApJ (2002) Sep. 20 issue, in pres

Two-Dimensional MHD Numerical Simulations of Magnetic Reconnection Triggered by A Supernova Shock in Interstellar Medium, Generation of X-Ray Gas in Galaxy

We examine the magnetic reconnection triggered by a supernova (or a point explosion) in interstellar medium, by performing two-dimensional resistive magnetohydrodynamic (MHD) numerical simulations with high spatial resolution. We found that the magnetic reconnection starts long after a supernova shock (fast-mode MHD shock) passes a current sheet. The current sheet evolves as follows: (i) Tearing-mode instability is excited by the supernova shock, and the current sheet becomes thin in its nonlinear stage. (ii) The current-sheet thinning is saturated when the current-sheet thickness becomes comparable to that of Sweet-Parker current sheet. After that, Sweet-Parker type reconnection starts, and the current-sheet length increases. (iii) ``Secondary tearing-mode instability'' occurs in the thin Sweet-Parker current sheet. (iv) As a result, further current-sheet thinning occurs and anomalous resistivity sets in, because gas density decreases in the current sheet. Petschek type reconnection starts and heats interstellar gas. Magnetic energy is released quickly while magnetic islands are moving in the current sheet during Petschek type reconnection. The released magnetic energy is determined by the interstellar magnetic field strength, not energy of initial explosion nor distance to explosion. We suggest that magnetic reconnection is a possible mechanism to generate X-ray gas in Galaxy.Comment: 17 pages using emulateapj.sty, 24 figures (4colors), submitted to ApJ, mpeg simulations and psfiles are available at http://stesun8.stelab.nagoya-u.ac.jp/~tanuma/apj2000/apj2000.htm

TRANS-ACTIVATION OF THE HUMAN SOX3 PROMOTER BY MAZ IN NT2/D1 CELLS

Abstract — In this study, we examine the role of three highly conserved putative binding sites for Myc-associated zinc finger protein (MAZ) in regulation of the human SOX3 gene expression. Electrophoretic mobility shift and supershift assays indicate that complexes formed at two out of three MAZ sites of the human SOX3 promoter involve ubiquitously expressed MAZ protein. Furthermore, in cotransfection experiments we demonstrate that MAZ acts as a positive regulator of SOX3 gene transcription in both undifferentiated and RA-differentiated NT2/D1 cells. Although MAZ increased both basal and RA-induced promoter activity, our results suggest that MAZ does not contribute to RA inducibility of the SOX3 promoter during neuronal differentiation of NT2/D1 cells

Molecular cloning and characterization of CIDE-3, a novel member of the cell-death-inducing DNA-fragmentation-factor (DFF45)-like effector family.

DNA fragmentation is one of the critical steps in apoptosis, which is induced by DNA fragmentation factor (DFF). DFF is composed of two subunits, a 40 kDa caspase-activated nuclease (DFF40) and a 45 kDa inhibitor (DFF45). Recently a novel family of cell-death-inducing DFF45-like effectors (CIDEs) has been identified. Among CIDEs, two from human (CIDE-A and CIDE-B) and three from mouse (CIDE-A, CIDE-B and FSP27) have been reported. In this study human CIDE-3, a novel member of CIDEs, was identified upon sequence analysis of a previously unidentified cDNA that encoded a protein of 238 amino acids. It was shown to be a human homologue of mouse FSP27, and shared homology with the CIDE-N and CIDE-C domains of CIDEs. Apoptosis-inducing activity was clearly shown by DNA-fragmentation assay of the nuclear DNA of CIDE-3 transfected 293T cells. The expression pattern of CIDE-3 was different from that of CIDE-B. As shown by Northern-blot analysis, CIDE-3 was expressed mainly in human small intestine, heart, colon and stomach, while CIDE-B showed strong expression in liver and small intestine and at a lower level in colon, kidney and spleen. Green-fluorescent-protein-tagged CIDE-3 was revealed in some cytosolic corpuscles. Alternative splicing of the CIDE-3 gene was also identified by reverse transcription PCR, revealing that two transcripts, CIDE-3 and CIDE-3alpha, were present in HepG2 and A375 cells. CIDE-3 comprised a full-length open reading frame with 238 amino acids; in CIDE-3alpha exon 3 was deleted and it encoded a protein of 164 amino acids. Interestingly the CIDE-3alpha isoform still kept the apoptosis-inducing activity and showed the same pattern of subcellular localization as CIDE-3. Consistent with its chromosome localization at 3p25, a region associated with high frequency loss of heterozygosity in many tumours, CIDE-3 may play an important role in prevention of tumorigenesis