Molecular mechanism underlying cAMP-induced amplification of IGF mitogenic activity through PI3-kinase binding protein, PI3KAP/XB130.

• Project/Area Number: 21580345
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied animal science
• Research Institution: The University of Tokyo
• Principal Investigator: HAKUNO Fumihiko 東京大学, 大学院・農学生命科学研究科, 助教 (30282700)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2011: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2009: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: IGF-I / PI 3-kinase / PI3KAP / 細胞増殖 / PITKAP / PI3-kinase

Research Abstract

We previously demonstrated that long-term pretreatment of rat FRTL-5 thyroid cells with TSH or cAMP-generating reagents potentiated IGF-I-dependent DNA synthesis. Under this condition, cAMP treatment increased tyrosine phosphorylation of a 125 kDa protein(p125) and its association with a p85 regulatory subunit of phosphatidylinositol 3-kinase(p85 PI3K), which were suggested to be important for potentiation of DNA synthesis. This study was undertaken to identify p125 and to elucidate its roles in potentiation of DNA synthesis induced by IGF-I. Immunoprecipitated phosphotyrosyl p125 in cAMP-stimulated FRTL-5 cells, was shown by MALDI-TOF MS analysis, to be a rat orthologue of human XB130, which we named phosphatidylinositol 3-kinase-associated protein(PI3KAP). cAMP treatment elevated PI3KAP/XB130 mRNA and protein levels as well as tyrosine phosphorylation and PI3KAP/XB130 interaction with p85 PI3K, leading to increased PI3K activities. Importantly, PI3KAP/XB130 knockdown in FRTL-5 cells attenuated cAMP-dependent potentiation of IGF-I-induced DNA synthesis. Addition of Src family kinase inhibitors, PP1 or PP2, during cAMP treatment abolished tyrosine phosphorylation of PI3KAP/XB130 and its interaction with p85 PI3K. In addition, c-Src was associated with PI3KAP/XB130 and was activated in response to cAMP. Together, these data indicate that cAMP-dependent induction of PI3KAP/XB130 and its association with PI3K are required for enhancement of IGF mitogenic activities.

Research Products

• [Journal Article] phosphatidylinositol 3-kinase-binding protein, PI3KAP/XB130, is required for cAMP-induced amplification of IGF mitogenic activity in FRTL-5 thyroid cells (2012)
  • Author(s): Yamanaka D, Akama T, Fukushima T, Nedachi T, Kawasaki C, Chida K, Minami S, Suzuki K, Hakuno F, Takahashi
  • Journal Title: Mol. Endocrinology Volume: 26 Pages: 1043-1055
• [Journal Article] (2012)
  • Author(s): Nakae J, Cao Y, Hakuno F, Takemori H, Kawano Y, Sekioka R, Abe T, Kiyonari H, Tanaka T, Sakai J, Takahashi S, Itoh H.
  • Journal Title: EMBO J Volume: 31 Pages: 2275-95
• [Journal Article] Kanzaki Novel missense mutation in the IGF-I receptor L2 domain results in intrauterine and postnatal growth retardation Clin (2012)
  • Author(s): Kawashima Y, Higaki K, Fukushima T, Hakuno F, Nagaishi JI, Hanaki K, Nanba E, Takahashi SI
  • Journal Title: Endocrinol
• [Journal Article] Enhanced oxidative stress in GH-transgenic rat and acromegaly in humans (2012)
  • Author(s): Nishizawa H, Handayaningsih AE, Iguchi G, Cho Y, Takahashi M, Yamamoto M, Suda K, Kasahara K, Hakuno F, Yamanouchi K, Nishihara M, Seino S, Takahashi S
  • Journal Title: Growth Horm. IGF Res Volume: 22 Pages: 64-68
• [Journal Article] Novel missense mutation in the IGF-I receptor L2 domain results in intrauterine and postnatal growth retardation (2012)
  • Author(s): Kawashima Y, Higaki K, Fukushima T, Hakuno F, Nagaishi JI, Hanaki K, Nanba E, Takahashi SI, Kanzaki S
  • Journal Title: Clin.Endocrinol. Volume: (in press) Issue: 2 Pages: 246-54
  • DOI: 10.1111/j.1365-2265.2012.04357.x
  • Peer Reviewed
• [Journal Article] phosphatidylinositol 3-kinase-binding protein, PI3KAP/XB130, is required for cAMP-induced amplification of IGF mitogenic activity in FRTL-5 thyroid cells (2012)
  • Author(s): Yamanaka D, Akama T, Fukushima T, Nedachi T, Kawasaki C, Chida K, Minami S, Suzuki K, Hakuno F, Takahashi S
  • Journal Title: Mol.Endocrinology Volume: (in press)
  • Peer Reviewed
• [Journal Article] Enhanced oxidative stress in GH-transgenic rat and acromegaly in humans (2012)
  • Author(s): Nishizawa H, Handayaningsih AE, Iguchi G, Cho Y, Takahashi M, Yamamoto M, Suda K, Kasahara K, Hakuno F, Yamanouchi K, Nishihara M, Seino S, Takahashi S, Takahashi Y
  • Journal Title: Growth Horm.IGF Res. Volume: 22 Issue: 2 Pages: 64-68
  • DOI: 10.1016/j.ghir.2012.02.001
  • Peer Reviewed
• [Journal Article] Constitutive expression of insulin receptor substrate(IRS)-1 inhibits myogenic differentiation through nuclear exclusion of Foxo1 in L6 myoblasts (2011)
  • Author(s): Hakuno F, Yamauchi Y, Kaneko G, Yoneyama Y, Nakae J, Chida K, Kadowaki T, Yamanouchi K, Nishihara M, Takahashi S-I
  • Journal Title: PLos One Volume: 6
• [Journal Article] Insulin receptor substrates form high-molecular-mass complexes that modulate their availability to insulin/insulin-like growth factor-I receptor tyrosine kinases (2011)
  • Author(s): Fukushima T, Arai T, Ariga-Nedachi M, Okajima H, Ooi Y, Iijima Y, Sone M, Cho Y, Ando Y, Kasahara K, Ozoe A, Yoshihara H, Chida K, Okada S, Kopchick JJ, Asano T, Hakuno F, Takahashi S-I.
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 404 Pages: 767-773
• [Journal Article] KIBRA suppresses apical exocytosis through inhibition of aPKC kinase activity in epithelial cells (2011)
  • Author(s): Yoshihama Y, Sasaki K, Horikoshi Y, Suzuki A, Ohtsuka T, Hakuno F, Takahashi S-I, Ohno S, Chida K.
  • Journal Title: Curr. Biol Volume: 21 Pages: 705-711
• [Journal Article] PKC promotes a proliferation to differentiation switch in keratinocytes via upregulation of p27Kip1 mRNA through suppression of JNK/c-Jun signaling under stress conditions (2011)
  • Author(s): Hara T, Miyazaki M, Hakuno F, Takahashi S-I, Chida K.
  • Journal Title: Cell Death Dis Volume: 2
• [Journal Article] HSP90 interacting with IRS-2 is involved in camp-dependent potentiation of IGF-I signals in FRTL-5 cells (2011)
  • Author(s): Fukushima T, Okajima H, Yamanaka D, Ariga M, Nagata S, Ito A, Yoshida M, Asano T, Chida K, Hakuno F, Takahashi S-I.
  • Journal Title: Mol. Cell Endocrinol. Volume: 344 Pages: 81-89
• [Journal Article] KIBRA suppresses apical exocytosis through inhibition of aPKC activity in epithelial cells (2011)
  • Author(s): Yohei Yoshihama
  • Journal Title: Current Biology Volume: 21 Issue: 8 Pages: 705-711
  • DOI: 10.1016/j.cub.2011.03.029
  • Peer Reviewed
• [Journal Article] PKC I promotes a proliferation to differentiation switch in keratinocytes via upregulation of p27Kip1 mRNA through suppression of JNK/c-Jun signaling under stress conditions (2011)
  • Author(s): Hara T, Miyazaki M, Hakuno F, Takahashi S, Chida K
  • Journal Title: Cell Death Dis. Volume: 2 Issue: 5 Pages: e157-e157
  • DOI: 10.1038/cddis.2011.40
  • Peer Reviewed
• [Journal Article] HSP90 interacting with IRS-2 is involved in cAMP-dependent potentiation of IGF-I signals in FRTL-5 cells (2011)
  • Author(s): Fukushima T, Okajima H, Yamanaka D, Ariga M, Nagata S, Ito A, Yoshida H, Asano T, Chida K
  • Journal Title: Mol.Cell Endocrinol. Volume: 344 Issue: 1-2 Pages: 81-90
  • DOI: 10.1016/j.mce.2011.06.029
  • Peer Reviewed
• [Journal Article] Constitutive expression of insulin receptor substrate(IRS)-1 inhibits myogenic differentiation through nuclear exclusion of Foxo1 in L6 myoblasts (2011)
  • Author(s): Hakuno F, Yamauchi Y, Kaneko G, Yoneyama Y, Nakae J, Chida K, Kadowaki T, Yamanouchi K, Nishihara M, Takahashi S
  • Journal Title: PLoS ONE Volume: 6 Issue: 10 Pages: e25655-e25655
  • DOI: 10.1371/journal.pone.0025655
  • Peer Reviewed
• [Journal Article] Insulin receptor substrates form high-molecular-mass complexes that modulate their availability to insulin/insulin-like growth factor-I receptor tyrosine kinase. (2011)
  • Author(s): Fukushima T. Hakuno F, et al.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 404 Pages: 767-773
  • Peer Reviewed
• [Journal Article] Effect of paraquat-induced oxidative stress on insulin regulation of insulin-like growth factor-binding protein-1 gene expression (2010)
  • Author(s): Kimura K, Katsumata Y, Ozawa T, Tawara S, Igarashi K, Cho Y, Shibata N, Hakuno F, Takahashi S-I, Takenaka A.
  • Journal Title: J. Clin. Biochem. Nutr Volume: 46 Pages: 157-167
  • NAID: 130004466606
• [Journal Article] Insulin receptor substrate-3, interacting with Bcl-3, enhances p50 NF-kappaB activity (2010)
  • Author(s): Kabuta T, Hakuno F, Cho Y, Yamanaka D, Chida K, AsanoT, Wada K, Takahashi S-I.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 394 Pages: 697-702
• [Journal Article] Paraquat-induced oxidative stress represses phosphatidylinositol 3-kinase activities leading to impaired glucose uptake in 3T3-L1 adipocytes (2010)
  • Author(s): Shibata M, Hakuno F, Yamanaka D, Okajima H, Fukushima T, Hasegawa T, Ogata T, Toyoshima Y, Chida K, Kimura K, Sakoda H, Takenaka A, Asano T, Takahashi S-I.
  • Journal Title: J. Biol. Chem. Volume: 285 Pages: 20915-25
• [Journal Article] Dietary protein deprivation upregulates insulin signaling and inhibits gluconeogenesis in rat liver (2010)
  • Author(s): Toyoshima Y, Tokita R, Ohne Y, Hakuno F, Noguchi T, Minami S, Kato H, Takahashi S-I.
  • Journal Title: J. Mol. Endocrinol. Volume: 45 Pages: 329-340
• [Journal Article] Paraquat-induced oxidative stress represses phosphatidylinositol 3-kinase activities leading to impaired glucose uptake in 3T3-L1 adipocytes. (2010)
  • Author(s): Shibata M, Hakuno F, et al.
  • Journal Title: J. Biol. Chem. Volume: 285 Pages: 20915-20925
  • Peer Reviewed
• [Journal Article] Dietary protein deprivation upregulates insulin signaling and inhibits gluconeogenesis in rat liver. (2010)
  • Author(s): Toyoshima Y, Hakuno F, et al.
  • Journal Title: J. Mol. Endocrinol. Volume: 45 Pages: 329-340
  • Peer Reviewed
• [Journal Article] Insulin receptor substrate-3, interacting with Bcl-3, enhances p50 NF-kappaB activity (2010)
  • Author(s): Kabuta T, Hakuno F, Cho Y, Yamanaka D, Chida K, Asano T, Wada K, Takahashi SI
  • Journal Title: Biochem.Biophys.Res.Commun. (in press)
  • Peer Reviewed
• [Journal Article] Effect of paraquat-induced oxidative stress on insulin regulation of insulin-like growth factor-binding protein-1 gene expression (2010)
  • Author(s): Kimura K, Katsumata Y, Ozawa T, Tawara S, Igarashi K, Cho Y, Shibata N, Hakuno F, Takahashi SI, Takenaka A
  • Journal Title: J.Clin.Biochem.Nutr. 46 Pages: 157-167
  • NAID: 130004466606
  • Peer Reviewed
• [Journal Article] Growth hormone(GH) and Insulin-like growth factor(IGF)-I represses 11beta-hydroxysteroid dehydrogenase type 1(HSD1) mRNA expression in 3T3-L1 cells and its activity in their homogenates (2009)
  • Author(s): Morita J, Hakuno F, Hizuka N, Takahashi S-I, Takano K.
  • Journal Title: Endocrine J. Volume: 56 Pages: 561-570
• [Journal Article] GH or IGF-I represses 11beta-hydroxysteroid dehydrogenase type 1(HSD1)mRNA expression in 3T3-L1 and its activity in their homogenates. (2009)
  • Author(s): Morita J, Hakuno F, Hizuka N, Takahashi SI, Takano K
  • Journal Title: Endocrine Journal 56 Pages: 561-570
  • Peer Reviewed
• [Presentation] PI 3-kinase activation through c-Src-dependent tyrosine phosphorylation of PI3KAP, a novel binding partner of a PI 3-kinase p85 regulatory subunit, plays novel roles in priming cells to IGF (2010)
  • Author(s): Yamanaka D, Akama T, Fukushima T, Minami S, Chida K, Hakuno F, Takahashi SI.
  • Organizer: The 5th international congress of the GRS and IGF society
  • Place of Presentation: New York(USA)
  • Year and Date: 2010-10-04
• [Presentation] PI 3-kinase activation through c-Src-dependent tyrosine phosphorylation of PITKAP, anovel bining partner of a PI 3-kinase p85 regulatory subunit, plays novel roles in priming (2010)
  • Author(s): 山中大介、伯野史彦, ら
  • Organizer: The Fifth International Congress of the GRS and IGF Society
  • Place of Presentation: New York (USA)
  • Year and Date: 2010-10-04
• [Presentation] PI 3-kinase activity associated with PI3KAP/XB130 that is recruited to F-actin is required for potentiation of IGF-I-induced cell proliferation (2009)
  • Author(s): Yamanaka D, Akama T, Fukushima T, Minami S, Chida K, Hakuno F, Takahashi SI.
  • Organizer: 第32回分子生物学会年会
  • Year and Date: 2009-12-11
• [Presentation] PI 3-kinase activity associated with PITKAP/XB 130 that is recruited to F-actin is required for potentiation of IGF-I-induced cell proliferation. (2009)
  • Author(s): Yamanaka D, Akama T, Fukushima T, Minami S, Chida K, Hakuno F, Takahashi SI
  • Organizer: 第32回分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-12-11
• [Book] 発達と成長を司るホルモン、インスリン様成長因子 (2010)
  • Author(s): 高橋伸一郎、竹中麻子、豊島由香、木村久美、加藤久典、福嶋俊明、山中大介、伯野史彦、南史朗、岩下光利
  • Publisher: 日本未熟児新生児学会雑誌
• [Remarks]
  • URL: http://endo.ar.a.u-tokyo.ac.jp/lab/shingroup/index.html
• [Remarks]
  • URL: http://endo.ar.a.u-tokyo.ac.jp/lab/index.html