2001 Fiscal Year Annual Research Report

褐色脂肪組織のエネルギー代謝における分子シンクロナイゼーションとそのメカニズム

Research Project

Project/Area Number	13022247
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (A)
Research Institution	The University of Tokushima
Principal Investigator	篠原康雄徳島大学, 薬学部, 助教授 (60226157)
Keywords	褐色脂肪組織 / エネルギー代謝 / 分子シンクロナイゼーション / H-FABP / 脂肪酸結合タンパク質 / 協調的転写制御
Research Abstract	本研究では、1.褐色脂肪組織のエネルギー代謝における分子シンクロナイゼーションを明らかにすることによって、この組織における適応型熱産生に必要なデバイスを分子レベルで理解すること、ならびに2.分子シンクロナイゼーションを制御する因子を同定し、生体内での協調的な遺伝子発現制御機構の様式を分子レベルで理解するとともに、これを新規のデバイス創成のための糸口とすることの2点を主たる目標とした実験を行う。初年度には、申請者らが最近見いだした、褐色脂肪組織の機能亢進に伴ったUCP1とH-FABPの協調的な転写亢進に焦点をあて、その分子機構を理解することを目的とした実験を行い、以下の知見を得た。a)H-FABP遺伝子の転写調節領域をクローン化し、本遺伝子の褐色脂肪組織での転写開始部位の決定を行った。その結果、褐色脂肪組織でも、筋組織と同じ領域で転写が始まることが判明、alternative promoterの存在の可能性が否定された。b)申請者らが従来用いてきた褐色脂肪細胞の初代培養系のcharacte rizationを行い、これがレチノイン酸やノルエピネフリンなど、vivoでの褐色脂肪組織の機能亢進に密接に関連しているとされている多くの刺激に適切に応答することから、転写制御機構の解析に有用であることを明らかにした。そこで、c)H-FABP遺伝子の転写調節領域を含むレポータープラスミドを調製し、これを初代培養した褐色脂肪細胞に導入、転写調節能の解析を行った。その結果、転写開始部位に隣接したproximal promoter regionが、レチノイン酸への応答に重要であることが明らかになった。

Research Products

(14 results)

All Other

All Publications (14 results)

[Publications] Y.Shinohara et al.: "Growth condition dependent synchronized changes in the transcript levels of type II hexokinase and type 1 glucose transporter in tumor cells"Biochim. Biophys. Acta. 1499. 242-248 (2001)
[Publications] S.Shimizu et al.: "Essential role of voltage-dependent anion channel in various forms of apoptosis in mammalian cells"J.Cell Biol.. 152. 237-250 (2001)
[Publications] Y.Shinohara et al.: "Expression of NAD+-dependent isocitrate dehydrogenase in brown adipose tissue"Biochem. Biophys. Res. Commun.. 281. 634-638 (2001)
[Publications] E.Majima et al.: "Specific labeling of the bovine heart mitochondrial phosphate carrier with fluorescein-5-isothiocyanate : roles of Lys185 and putative adenine nucleotide recognition site in phosphate transport"J.Biol.Chem.. 276. 9792-9799 (2001)
[Publications] T.Hatanaka et al.: "Significant expression of functional human type 1 mitochondrial ADP/ATP carrier in yeast mitochondria"Biol. Pharm. Bull.. 24. 595-599 (2001)
[Publications] T.Hatanaka et al.: "Significant effect of the N-terminal region of the yeast type 2 mitochondrial ADP/ATP carrier on efficient expression in yeast mitochondria"J.Biol.Chem.. 276. 28881-28888 (2001)
[Publications] T.Hatanaka et al.: "Characterization of loops of the yeast mitochondrial ADP/ATP carrier facing the cytosol by site-directed mutagenesis"Biochem. Biophys. Res. Commun.. 286. 936-942 (2001)
[Publications] H.Harashima et al.: "Intracellular control of gene trafficking using liposome as drug carriers"Eur. J. Pharm. Sci.. 13. 85-89 (2001)
[Publications] R.Tachibana et al.: "Intracellular control of macromolecules by use of liposome as drug carriers"J.Biol.Macromol.. 1. 60-67 (2001)
[Publications] R.Tachibana et al.: "Quantitative studies on the nuclear transport of plasmid DNA and gene expression employing nonviral vectors"Advanced Drug Delivery Reviews. 52. 219-226 (2001)
[Publications] M.Hino et al.: "Requirement of continuous transcription for production of certain amount of protein by Rapid Translation System"Protein Express. Purif.. (in press).
[Publications] N.Yamazaki et al.: "Structural properties of mammalian mitochondrial ADP/ATP carriers"Mitochondrion. (in press).
[Publications] M.Tabuchi et al.: "Cell-free protein synthesis on a microchip"Proteomics. (in press).
[Publications] R.Tachibana et al.: "Relationship between the amount of plasmid delivered into nucleus and gene expression revealed by quantitative evaluation of intranuclear plasmid"Pharm. Res.. (in press).

2001 Fiscal Year Annual Research Report

褐色脂肪組織のエネルギー代謝における分子シンクロナイゼーションとそのメカニズム

Principal Investigator

篠原 康雄 徳島大学, 薬学部, 助教授 (60226157)

Research Products

[Publications] Y.Shinohara et al.: "Growth condition dependent synchronized changes in the transcript levels of type II hexokinase and type 1 glucose transporter in tumor cells"Biochim. Biophys. Acta. 1499. 242-248 (2001)

[Publications] S.Shimizu et al.: "Essential role of voltage-dependent anion channel in various forms of apoptosis in mammalian cells"J.Cell Biol.. 152. 237-250 (2001)

[Publications] Y.Shinohara et al.: "Expression of NAD+-dependent isocitrate dehydrogenase in brown adipose tissue"Biochem. Biophys. Res. Commun.. 281. 634-638 (2001)

[Publications] E.Majima et al.: "Specific labeling of the bovine heart mitochondrial phosphate carrier with fluorescein-5-isothiocyanate : roles of Lys185 and putative adenine nucleotide recognition site in phosphate transport"J.Biol.Chem.. 276. 9792-9799 (2001)

[Publications] T.Hatanaka et al.: "Significant expression of functional human type 1 mitochondrial ADP/ATP carrier in yeast mitochondria"Biol. Pharm. Bull.. 24. 595-599 (2001)

[Publications] T.Hatanaka et al.: "Significant effect of the N-terminal region of the yeast type 2 mitochondrial ADP/ATP carrier on efficient expression in yeast mitochondria"J.Biol.Chem.. 276. 28881-28888 (2001)

[Publications] T.Hatanaka et al.: "Characterization of loops of the yeast mitochondrial ADP/ATP carrier facing the cytosol by site-directed mutagenesis"Biochem. Biophys. Res. Commun.. 286. 936-942 (2001)

[Publications] H.Harashima et al.: "Intracellular control of gene trafficking using liposome as drug carriers"Eur. J. Pharm. Sci.. 13. 85-89 (2001)

[Publications] R.Tachibana et al.: "Intracellular control of macromolecules by use of liposome as drug carriers"J.Biol.Macromol.. 1. 60-67 (2001)

[Publications] R.Tachibana et al.: "Quantitative studies on the nuclear transport of plasmid DNA and gene expression employing nonviral vectors"Advanced Drug Delivery Reviews. 52. 219-226 (2001)

[Publications] M.Hino et al.: "Requirement of continuous transcription for production of certain amount of protein by Rapid Translation System"Protein Express. Purif.. (in press).

[Publications] N.Yamazaki et al.: "Structural properties of mammalian mitochondrial ADP/ATP carriers"Mitochondrion. (in press).

[Publications] M.Tabuchi et al.: "Cell-free protein synthesis on a microchip"Proteomics. (in press).

[Publications] R.Tachibana et al.: "Relationship between the amount of plasmid delivered into nucleus and gene expression revealed by quantitative evaluation of intranuclear plasmid"Pharm. Res.. (in press).

篠原康雄徳島大学, 薬学部, 助教授 (60226157)