2002 Fiscal Year Annual Research Report

細胞極性確立とmRNA局在のin vivo イメージング

Research Project

Project/Area Number	11242202
Research Institution	Osaka University
Principal Investigator	入江賢児大阪大学, 医学系研究科, 助教授 (90232628)
Keywords	非対称分裂 / mRNA / RNA結合蛋白質 / 酵母 / 細胞骨格 / アクチン / 翻訳 / ミオシン
Research Abstract	本研究では、細胞極性の分子レベルでの研究が最も進んでいる出芽酵母を用いて、mRNAや細胞極性決定因子の局在のin vivoイメージングの系を確立し、その系を用いてmRNAの細胞内局在の分子機構機構および細胞極性の確立や維持に関与する因子のmRNA局在における機能を明らかにする。また、動物細胞から酵母相同遺伝子を機能的に分離することにより、動物細胞におけるmRNAの局在機構およびその細胞極性との関連を明らかにする。本年度は、酵母および培養細胞を用いた細胞極性、非対称分裂、mRNAの局在機構の解析から、以下の結果を得た。 (1)細胞分裂時におけるASH1 mRNAの娘細胞の先端への局在を、生きた酵母細胞中で観察する系を用いて、ASH1 mRNAと共局在するRNA結合タンパク質Khd1を見出した。 (2)Khd1がASH1 mRNAのコーディング配列内の領域に結合し、その翻訳を抑制することを明らかにした。 (3)PufファミリーのRNA結合モチーフをもつMpt5が、HO mRNAの3'非翻訳領域に結合し、HO遺伝子の発現を抑制することを見出した。 (4)NucleaseドメインをもつMkt1が、HO mRNAの3'非翻訳領域を介して、HO遺伝子の発現を正に制御することを明らかにした。 (5)上皮細胞におけるタイトジャンクションの接着分子JAMの局在が、ネクチン-アファディン系の細胞間接着系によって決定されることを明らかにした。このように、本年度の研究は予想以上に進展し、当初の目的を達成することができた。

Research Products

(12 results)

All Other

All Publications (12 results)

[Publications] Irie, K.: "The Khd1 protein, which has three KH RNA-binding motifs, is required for proper localization of ASH1 mRNA in yeast"EMBO J.. 21・5. 58-67 (2002)
[Publications] Ozaki-Kuroda, K.: "Nectin couples cell-cell adhesion and the actin scaffold at heterotypic testicular junctions"Curr. Biol.. 12・13. 1145-1150 (2002)
[Publications] Fukuhara, A.: "Involvement of nectin in the localization of junctional adhesion molecule at tight junctions"Oncogene. 21・50. 7642-7655 (2002)
[Publications] Momose, Y.: "Role of the second immunoglobulin-like loop of nectin in cell-cell adhesion"Biochem. Biophys. Res. Commun.. 293・1. 45-49 (2002)
[Publications] Tanaka, Y.: "Ectodomain shedding of nectin-1α by SF/HGF and TPA in MDCK cells"Biochem. Biophys. Res. Commun.. 299・3. 472-478 (2002)
[Publications] Fukuhara, A.: "Role of nectin in organization of tight junctions in epithelial cells"Genes Cells. 7・10. 1059-1072 (2002)
[Publications] Asada, M.: "ADIP, a novel afadin-and α-actinin-binding protein localized at cell-cell adherens junctions"J. Biol. Chem.. 278・6. 4103-4111 (2003)
[Publications] Honda, T.: "Antagonistic and agonistic effects of an extracellular fragment of nectin on formation of E-cadherin-based cell-cell adhesion"Genes Cells. 8・1. 51-63 (2003)
[Publications] Toi, H.: "She4p/Dim1p interacts with the motor domain of unconventional myosins in the budding yeast, Saccharomyces cerevisiae"Mol. Biol. Cell. (in press). (2003)
[Publications] Katata, T.: "Involvement of nectin in the localization of IQGAP1 at the cell-cell adhesion sites through the actin cytoskeleton in Madin-Darby canine kidney cells"Oncogene. (in press). (2003)
[Publications] Honda, T.: "Cdc42 and Rac small G proteins activated by trans-interactions of nectins are involved in activation of c-Jun N-terminal kinase, but not in association of nectins and E-cadherin to form adherens junctions, in fibroblasts"Genes Cells. (in press). (2003)
[Publications] Inagaki, M.: "Nectin-dependent localization of ZO-1 at puncta adhaerentia junctions between the mossy fiber terminals and the dendrites of the pyramidal cells in the CA3 area of adult mouse hippocampus"J. Comp. Neurol.. (in press). (2003)

2002 Fiscal Year Annual Research Report

細胞極性確立とmRNA局在のin vivo イメージング

Principal Investigator

入江 賢児 大阪大学, 医学系研究科, 助教授 (90232628)

Research Products

[Publications] Irie, K.: "The Khd1 protein, which has three KH RNA-binding motifs, is required for proper localization of ASH1 mRNA in yeast"EMBO J.. 21・5. 58-67 (2002)

[Publications] Ozaki-Kuroda, K.: "Nectin couples cell-cell adhesion and the actin scaffold at heterotypic testicular junctions"Curr. Biol.. 12・13. 1145-1150 (2002)

[Publications] Fukuhara, A.: "Involvement of nectin in the localization of junctional adhesion molecule at tight junctions"Oncogene. 21・50. 7642-7655 (2002)

[Publications] Momose, Y.: "Role of the second immunoglobulin-like loop of nectin in cell-cell adhesion"Biochem. Biophys. Res. Commun.. 293・1. 45-49 (2002)

[Publications] Tanaka, Y.: "Ectodomain shedding of nectin-1α by SF/HGF and TPA in MDCK cells"Biochem. Biophys. Res. Commun.. 299・3. 472-478 (2002)

[Publications] Fukuhara, A.: "Role of nectin in organization of tight junctions in epithelial cells"Genes Cells. 7・10. 1059-1072 (2002)

[Publications] Asada, M.: "ADIP, a novel afadin-and α-actinin-binding protein localized at cell-cell adherens junctions"J. Biol. Chem.. 278・6. 4103-4111 (2003)

[Publications] Honda, T.: "Antagonistic and agonistic effects of an extracellular fragment of nectin on formation of E-cadherin-based cell-cell adhesion"Genes Cells. 8・1. 51-63 (2003)

[Publications] Toi, H.: "She4p/Dim1p interacts with the motor domain of unconventional myosins in the budding yeast, Saccharomyces cerevisiae"Mol. Biol. Cell. (in press). (2003)

[Publications] Katata, T.: "Involvement of nectin in the localization of IQGAP1 at the cell-cell adhesion sites through the actin cytoskeleton in Madin-Darby canine kidney cells"Oncogene. (in press). (2003)

[Publications] Honda, T.: "Cdc42 and Rac small G proteins activated by trans-interactions of nectins are involved in activation of c-Jun N-terminal kinase, but not in association of nectins and E-cadherin to form adherens junctions, in fibroblasts"Genes Cells. (in press). (2003)

[Publications] Inagaki, M.: "Nectin-dependent localization of ZO-1 at puncta adhaerentia junctions between the mossy fiber terminals and the dendrites of the pyramidal cells in the CA3 area of adult mouse hippocampus"J. Comp. Neurol.. (in press). (2003)

入江賢児大阪大学, 医学系研究科, 助教授 (90232628)