Cooperative contributions of Interferon regulatory factor 1 (IRF1) and IRF8 to interferon-γ-mediated cytotoxic effects on oligodendroglial progenitor cells

<p>Abstract</p> <p>Background</p> <p>Administration of exogenous interferon-γ (IFNγ) aggravates the symptoms of multiple sclerosis (MS), whereas interferon-β (IFNβ) is used for treatment of MS patients. We previously demonstrated that IFNγ induces apoptosis of oligodendroglial progenitor cells (OPCs), suggesting that IFNγ is more toxic to OPCs than IFNβ. Thus we hypothesized that a difference in expression profiles between IFNγ-inducible and IFNβ-inducible genes in OPCs would predict the genes responsible for IFNγ-mediated cytotoxic effects on OPCs. We have tested this hypothesis particularly focusing on the interferon regulatory factors (IRFs) well-known transcription factors up-regulated by IFNs.</p> <p>Methods</p> <p>Highly pure primary rat OPC cultures were treated with IFNγ and IFNβ. Cell death and proliferation were assessed by MTT reduction, caspse-3-like proteinase activity, Annexin-V binding, mitochondrial membrane potential, and BrdU-incorporation. Induction of all nine IRFs was comprehensively compared by quantitative PCR between IFNγ-treated and IFNβ-treated OPCs. IRFs more strongly induced by IFNγ than by IFNβ were selected, and tested for their ability to induce OPC apoptosis by overexpression and by inhibition by dominant-negative proteins or small interference RNA either in the presence or absence of IFNγ.</p> <p>Results</p> <p>Unlike IFNγ, IFNβ did not induce apoptosis of OPCs. Among nine IRFs, IRF1 and IRF8 were preferentially up-regulated by IFNγ. In contrast, IRF7 was more robustly induced by IFNβ than by IFNγ. Overexpressed IRF1 elicited apoptosis of OPCs, and a dominant negative IRF1 protein partially protected OPCs from IFNγ-induced apoptosis, indicating a substantial contribution of IRF1 to IFNγ-induced OPC apoptosis. On the other hand, overexpression of IRF8 itself had only marginal proapoptotic effects. However, overexpressed IRF8 enhanced the IFNγ-induced cytotoxicity and the proapoptotic effect of overexpressed IRF1, and down-regulation of IRF8 by siRNA partially but significantly reduced preapoptotic cells after treatment with IFNγ, suggesting that IRF8 cooperatively enhances IFNγ-induced OPC apoptosis.</p> <p>Conclusions</p> <p>This study has identified that IRF1 and IRF8 mediate IFNγ-signaling leading to OPC apoptosis. Therapies targeting at these transcription factors and their target genes could reduce IFNγ-induced OPC loss and thereby enhance remyelination in MS patients.</p

PROTEIN: A Visual Interface for Classification of Partial Reliefs

ABSTRACT 3D structure of proteins deeply takes part in the expression of the protein. Molecular surfaces of the protein generally have very complex and bumpy shapes. It is well-known that functions of proteins strongly appear in the bumpy parts of the molecular surfaces. We propose a visual interface to effectively visualize the partial reliefs of molecular surfaces of proteins. This technique assumes that molecule surfaces are approximated as triangular meshes. It first extracts groups of triangles forming partial reliefs, and calculates their feature values as histograms. It finally clusters the partial reliefs according to the histogram. The presented technique then visualizes the clustering results applying a hierarchical data visualization technique &quot;HeiankyoView&quot;, as a visual interface to explore the clustered partial reliefs

Near-Infrared Coronagraphic Observations of the T Tauri Binary System UY Aur

We present a near-infrared image of UY Aur, a 0.9" separated binary system, using the Coronagraphic Imager with Adaptive Optics on the Subaru Telescope. Thanks to adaptive optics, the spatial resolution of our image was ~0.1" in the full width at half maximum of the point spread function, the highest achieved. By comparison with previous measurements, we estimated that the orbital period is ~1640 yrs and the total mass of the binary is ~1.73 solar mass. The observed H-band magnitude of the secondary varies by as much as 1.3 mag within a decade, while that of the primary is rather stable. This inconstancy may arise from photospheric variability caused by an uneven accretion rate or from the rotation of the secondary. We detected a half-ring shaped circumbinary disk around the binary with a bright southwest part but a barely detectable northeast portion. The brightness ratio is ~57. Its inner radius and inclination are about 520 AU and 42, respectively. The disk is not uniform but has remarkable features, including a clumpy structure along the disk, circumstellar material inside the inner cavity, and an extended armlike structure. The circumstellar material inside the cavity probably corresponds to a clump or material accreting from the disk onto the binary. The armlike structure is a part of the disk, created by the accretion from the outer region of the disk or encounters with other stellar systems.Comment: 16 pages, 6 figures; accepted for publication in A

Contributions of Three Starch Branching Enzyme Isozymes to the Fine Structure of Amylopectin in Rice Endosperm

Three starch branching enzyme (BE) isozymes, BEI, BEIIa, and BEIIb, are involved in starch biosynthesis in rice endosperm. Past in vivo and in vitro studies have suggested that each BE isozyme plays a distinct role in forming the fine structure of amylopectin. To elucidate more details of their roles, we prepared DNA constructs in which all the possible combinations of the expressions of these three isozymes were suppressed in developing rice endosperm. Analysis of the chain-length distributions of amylopectin produced under these various conditions confirmed the contributions of the individual BE isozymes to the fine structure of amylopectin in rice endosperm. Among these isozymes, the impact of loss of BEIIb activity on amylopectin fine structure was most remarkable and indicated that it plays a specific role in the synthesis of short chains with a 6–13 degree of polymerization (DP). The contribution of BEI to the amylopectin synthesis was unclear when only BEI activity was reduced. It was clear, however, when both BEI and BEIIb activities were substantially inhibited. The DP11-22 intermediate chains were markedly reduced in the ΔBEI/BEIIb line compared with the ΔBEIIb line, indicating that BEI plays a distinct role in the synthesis of these intermediate chains. Although no substantial change in amylopectin chain profile was detected in the ΔBEIIa line, the role of BEIIa could be deciphered by analyzing amylopectin fine structure from the ΔBEI/BEIIa/BEIIb line in comparison to that from ΔBEI/BEIIb line. This strongly suggests that BEIIa compensates for the role of BEI, rather than that of BEIIb, by forming intermediate chains of DP11-22. In addition, the new possibility that BEIIa is involved in the formation of starch granules in rice endosperm was suggested because the onset temperature for gelatinization of starch granules in the ΔBEIIa/BEIIb line was significantly higher than that in the ΔBEIIb line. In summary, the present study highlights the distinct roles of BEI, BEIIa, and BEIIb in the synthesis of amylopectin in developing rice endosperm

Coupling of Rotation and Catalysis in F1-ATPase Revealed by Single-Molecule Imaging and Manipulation

SummaryF1-ATPase is a rotary molecular motor that proceeds in 120° steps, each driven by ATP hydrolysis. How the chemical reactions that occur in three catalytic sites are coupled to mechanical rotation is the central question. Here, we show by high-speed imaging of rotation in single molecules of F1 that phosphate release drives the last 40° of the 120° step, and that the 40° rotation accompanies reduction of the affinity for phosphate. We also show, by single-molecule imaging of a fluorescent ATP analog Cy3-ATP while F1 is forced to rotate slowly, that release of Cy3-ADP occurs at ∼240° after it is bound as Cy3-ATP at 0°. This and other results suggest that the affinity for ADP also decreases with rotation, and thus ADP release contributes part of energy for rotation. Together with previous results, the coupling scheme is now basically complete