2000 Fiscal Year Annual Research Report

マウス初期胚母性RNAの時期特異的ポリ(A)鎖伸長の意義

Research Project

Project/Area Number	12029221
Research Institution	Tokai University
Principal Investigator	木村穣東海大学, 医学部, 教授 (10146706)
Co-Investigator(Kenkyū-buntansha)	佐藤正宏東海大学, 総合医学研究所, 助教授 (30287099) 桜井敬之東海大学, 医学部, 助手 (80317825)
Keywords	マウス / 着床前初期胚 / 卵形成 / 受精卵 / 母性RNA / 接合型遺伝子発現 / poly(A)化 / 翻訳制御
Research Abstract	受精前後で発現する遺伝子群の研究の過程で単離した新規遺伝子のSSEC-D cDNA(stage specific embryonic clone-D)の解析から、その母性mRNAは1細胞期後期(受精後18時間)をピークとして一過的にmRNAのポリA鎖を伸長させていることと、この伸長には未知のcis配列が利用されていることを見いだした。マウスでは受精後ポリ(A)鎖を伸長する母性RNAの報告は無い。本研究の目的は、我々が見いだした受精後に生じる母性RNAのポリ(A)鎖伸長現象が、マウス初期胚発生プログラムにいかに関与するかを解明することである。本年度、この伸長に関わる未知のcis配列の同定に向け、EGFPcDNAおよび各鎖長のポリ(A)を持つ一連の融合SSEC-DRNAをマウス受精卵細胞質へ注入する実験を行い、幾つかの知見を得た。先ず、注入RNAは受精卵に本来存在するSSEC-D母性RNAと同様にRNA伸長を示し、注入実験系をほぼ確立できた。この系を駆使し、受精後に生じる母性RNAのポリA鎖伸長現象には、(i)SSEC-D配列および45ntds以上の3′末端のpoly(A)鎖の存在が必要であること。(ii)特定のSSEC-D配列の欠損したRNA注入実験から、SSEC-DのRNA配列中の特定の配列に依存していることを示した。さらに、3末端のpoly(A)鎖とSSEC-DのRNA配列中の特定の配列がRNA安定性に関与することを見いだし、EGFPの注入胚での発現から、3′末端のpoly(A)鎖の存在、もしくはpoly(A)鎖の伸長が翻訳活性への影響する可能性が認められた。これらの一連の結果から、ダイナミックな変動を示す母性型SSECDmRNAの動態に関するRNAシスシグナルおよびトランス因子の同定のための研究へ進展させるための足がかりが出来たと考える。

Research Products
(12 results)

All Other

All Publications (12 results)

[Publications] Sadahiro,S., et al: "Morphometric Analysis of the Myelin-Associated Oligodendrocyte Basic Protein-Deficient Mouse Reveals a Possible Role for Myelin-Associated Oligodendrocytis Basic Protein in Reguiating Axonal Diameter"Neuroscience. 98. 361-367 (2000)
[Publications] Tomita,K., et al.: "Gene structure and promoter for Crad2 encoding mouse cis-retinol/3alpfa-hydroxysterol short-chain dehydrogenase isozyme."Gene. 251. 175-186 (2000)
[Publications] Sato,M. et al.: "New Approach to Cell Lineage Analysis in Mammals Using Cre-loxP System."Molecular Reproduction and Fertility. 56. 34-44 (2000)
[Publications] Sato,M., et al.: "Testis-mediated gene transfer (TMGT) in mice : Attempts to improve TMGT using commercially available reagents used for gene transfer in mammalian culture system"Transgenics (2001). (in press).
[Publications] Kajiwara,K., et al.: "Sez4 gene encoding an elongation subunit of DNA polymerase zeta is required for normal embyogenesis"Genes to Cells (2001). (in press).
[Publications] Kawamura,K., et al.: "The error-prone DNA polymerase zeta catalytic subunit( Rev3) gene is ubiquitously expressed in normal and malignant human tissues."Int.J.Oncology. 18. 97-103 (2001)
[Publications] M.Shoji, et al.: "Age-related amyloid b protein accumulation induces cellular death and macrophage activation in transgenic mice."J.Pathol.. 191. 93-101 (2000)
[Publications] M.Sato, et al.: "Enhanced green fluorescent prote (EGFP) as a useful tag for identification of homozygous transgenic embryos."J.Reprod.Engineer.. 3. 124-133 (2000)
[Publications] M.Sato,T. et al.: "Enhanced green fluorescent protein as a useful tag for rapid identification of homozygous transgenic mice."Genetic Anal : Biomol. Engineer.(2000). (in press).
[Publications] M.Sato and M.Kimura: "Intrabursal transfer of spermatozoa (ITS) : A new route for artificial insemination of mice."Theriogenol.(2000). (in press).
[Publications] M.Sato,K.Miyado and M.Kimura: "Cloning and characterization of 5'-upstream sequence of the M32 gene for a mouse homologue of Drosophila heterochromatin protein 1 (HP1)."DNA Sequence (2000). (in press).
[Publications] Huang Z.Tamura M.Sakurai T et al: "In vivo transfection of testicular germ cells and transgenesis by using the mitochondrially localized jellyfish fluorescent protein gene."FEBS Lett.. 487(2). 248-251 (2000)

2000 Fiscal Year Annual Research Report

マウス初期胚母性RNAの時期特異的ポリ(A)鎖伸長の意義

Principal Investigator

木村 穣 東海大学, 医学部, 教授 (10146706)

Research Products

[Publications] Sadahiro,S., et al: "Morphometric Analysis of the Myelin-Associated Oligodendrocyte Basic Protein-Deficient Mouse Reveals a Possible Role for Myelin-Associated Oligodendrocytis Basic Protein in Reguiating Axonal Diameter"Neuroscience. 98. 361-367 (2000)

[Publications] Tomita,K., et al.: "Gene structure and promoter for Crad2 encoding mouse cis-retinol/3alpfa-hydroxysterol short-chain dehydrogenase isozyme."Gene. 251. 175-186 (2000)

[Publications] Sato,M. et al.: "New Approach to Cell Lineage Analysis in Mammals Using Cre-loxP System."Molecular Reproduction and Fertility. 56. 34-44 (2000)

[Publications] Sato,M., et al.: "Testis-mediated gene transfer (TMGT) in mice : Attempts to improve TMGT using commercially available reagents used for gene transfer in mammalian culture system"Transgenics (2001). (in press).

[Publications] Kajiwara,K., et al.: "Sez4 gene encoding an elongation subunit of DNA polymerase zeta is required for normal embyogenesis"Genes to Cells (2001). (in press).

[Publications] Kawamura,K., et al.: "The error-prone DNA polymerase zeta catalytic subunit( Rev3) gene is ubiquitously expressed in normal and malignant human tissues."Int.J.Oncology. 18. 97-103 (2001)

[Publications] M.Shoji, et al.: "Age-related amyloid b protein accumulation induces cellular death and macrophage activation in transgenic mice."J.Pathol.. 191. 93-101 (2000)

[Publications] M.Sato, et al.: "Enhanced green fluorescent prote (EGFP) as a useful tag for identification of homozygous transgenic embryos."J.Reprod.Engineer.. 3. 124-133 (2000)

[Publications] M.Sato,T. et al.: "Enhanced green fluorescent protein as a useful tag for rapid identification of homozygous transgenic mice."Genetic Anal : Biomol. Engineer.(2000). (in press).

[Publications] M.Sato and M.Kimura: "Intrabursal transfer of spermatozoa (ITS) : A new route for artificial insemination of mice."Theriogenol.(2000). (in press).

[Publications] M.Sato,K.Miyado and M.Kimura: "Cloning and characterization of 5'-upstream sequence of the M32 gene for a mouse homologue of Drosophila heterochromatin protein 1 (HP1)."DNA Sequence (2000). (in press).

[Publications] Huang Z.Tamura M.Sakurai T et al: "In vivo transfection of testicular germ cells and transgenesis by using the mitochondrially localized jellyfish fluorescent protein gene."FEBS Lett.. 487(2). 248-251 (2000)

木村穣東海大学, 医学部, 教授 (10146706)