SKELETAL MUSCLE SYNTROPHIN INTERACTORS REVEALED BY YEAST TWO-HYBRID ASSAY

2008-08Syntrophins are the cytoplasmic peripheral proteins of dystrophin glycoprotein complex, of which five (α1, β1, β2, γ1 and γ2) isoforms have been identified so far. Respective syntrophin isoforms are encoded by different genes but have similar domain structures. At the sarcolemma of skeletal muscle, the most abundant α1-syntrophin was shown to interact at its PDZ domain with many membrane proteins. Among them, the AQP4 interaction with α1-syntrophin PDZ domain was demonstrated by a Tg mouse study, prompting us to investigate the interaction between mouse α1-syntrophin (BC018546: nt.267–492, PDZ domain) pEXP-AD502 as prey vector and mouse AQP4 (NM009700: nt.805–969) pDBLeu as bait vector by the yeast two-hybrid assay, resulting in a negative study. We further studied the binding partner of another sarcolemma located β1-syntrophin, and performed a yeast two-hybrid experiment. With human β1-syntrophin as bait and human skeletal muscle cDNA library as prey, we obtained one positive clone which turned out to be α-dystrobrevin. Although the interaction of human β1-syntrophin with α-dystrobrevin has already been shown by immunoprecipitation assay, we have here confirmed this interaction by a yeast two-hybrid experiment.departmental bulletin pape

Studies of CP Violation in B→J/ψK* Decays

journal articl

アルカリ金属炭酸塩の熱力学状態図

application/pdfpostprintThe purpose of this work was to prepare the thermodynamical phase diagram which shows the stability of alkali carbonates under various environments. Several types of phase diagrams were discussed. Then, the log aC-log Po₂ diagram was proposed. The preparation procedures were described to show the stability region of alkali carbonates on the log aC-log Po₂ diagram. The diagram was compared with the well known E-pO²ˉ diagram. It was found that the E-pO²ˉ diagram is equivalent to log aM-log Po₂ diagram. Difficulty was emphasized in the use of E-pO²ˉ diagram for reducing atmospheres. On the other hand, the proposed log aC-log Po₂ diagram shows the stability of alkali carbonates by any two of the parameters aC, Po₂, Pco, Pco₂, and aM. It was shown that the EMF of the carbonate electrochemical cells are obtained from the pfoposed diagram.departmental bulletin pape

3P269 配列空間における構造自由エネルギー地形の描像(生命の起源・進化, ポスター発表, 第45回日本生物物理学会年会)

rights: 日本生物物理学会 rights: 本文データは学協会の許諾に基づきCiNiiから複製したものである relation: IsVersionOf: http://ci.nii.ac.jp/naid/110006563188/textapplication/pdfjournal articl

0812-9869-9940 (WA), Jual Keranda Mayat Panawaren

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Diphtheria toxin‐mediated transposon‐driven poly (A)‐trapping efficiently disrupts transcriptionally silent genes in embryonic stem cells

Random gene trapping is the application of insertional mutagenesis techniques that are conventionally used to inactivate protein‐coding genes in mouse embryonic stem (ES) cells. Transcriptionally silent genes are not effectively targeted by conventional random gene trapping techniques, thus we herein developed an unbiased poly (A) trap (UPATrap) method using a Tol2 transposon, which preferentially integrated into active genes rather than silent genes in ES cells. To achieve efficient trapping at transcriptionally silent genes using random insertional mutagenesis in ES cells, we generated a new diphtheria toxin (DT)‐mediated trapping vector, DTrap that removed cells, through the expression of DT that was induced by the promoter activity of the trapped genes, and selected trapped clones using the neomycin‐resistance gene of the vector. We found that a double‐DT, the dDT vector, dominantly induced the disruption of silent genes, but not active genes, and showed more stable integration in ES cells than the UPATrap vector. The dDT vector disrupted differentiated cell lineage genes, which were silent in ES cells, and labeled trapped clone cells by the expression of EGFP upon differentiation. Thus, the dDT vector provides a systematic approach to disrupt silent genes and examine the cellular functions of trapped genes in the differentiation of target cells and development.journal articl

Evaluation of Parameters Critical for Observing Nucleic Acids Inside Living <i>Xenopus laevis</i> Oocytes by In-Cell NMR Spectroscopy

In-cell NMR spectroscopy of proteins in different cellular environments is a well-established technique that, however, has not been applied to nucleic acids so far. Here, we show that isotopically labeled DNA and RNA can be observed inside the eukaryotic environment of Xenopus laevis oocytes by in-cell NMR spectroscopy. One limiting factor for the observation of nucleic acids in Xenopus oocytes is their reduced stability. We demonstrate that chemical modification of DNA and RNA can protect them from degradation and can significantly enhance their lifetime. Finally, we show that the imino region of the NMR spectrum is devoid of any oocyte background signals enabling the detection even of isotopically nonlabeled molecules