Molecular basis of inborn errors of ketone body metabolism : mainly basic studies for responsible genes

• Project/Area Number: 16591019
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Gifu University
• Principal Investigator: FUKAO Toshiyuki Gifu University, Graduate School of Medicine Department of Pediatrics, Associate professor, 大学院・医学系研究科, 助教授 (70260578)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: thiolase / CoA transferase / inborn errors of metabolism / ketone body metabolism / methylation / Alu sequence / "Mild" mutation / tertiary structure / nonsense-mediated RNA decay / T2 / SCOT / β-ケトチオラーゼ欠損症 / サクシニル-CoA:3-ケト酸CoAトランスフェラーゼ欠損症 / 遺伝子発現 / ノックアウトマウス / 3-ハイドロキシ酪酸脱水素酵素

Research Abstract

1.Succinyl-CoA:3-ketoacid CoA transferase (SCOT) deficiency
1)We revealed and proved that SCOT deficient patients with "mild" mutations did not always show permanent ketosis which is pathognomonic feature of SCOT deficiency.
2)In a SCOT deficient patient, we found exons 6 and 7 skipping, which was caused by G>A substitution at the end nucleotide of exon 6. We analyzed heteronuclear RNA and cytosolic RNA and revealed that mRNA with single exon 6 skipping was degraded rapidly by nonsense-mediated RNA decay. Since mRNA with exon 6 and 7 skipping was in frame and although it was minor transcript in nuclear RNA, this mRNA became a major product in cytosolic fraction.
2.Mitochondrial acetyoacetyl-CoA thiolase (T2) deficiency
1)A coupled assay with tiglyl-CoA had been widely used for the enzymatic diagnosis of T2 deficiency in Europe and USA. We revealed and proved that the result of coupled assay using tiglyl-CoA was normal in two T2 deficient patients with "mild" mutations. This means that T2 deficient patients with "mild" mutations had been mis-diagnosed as normal using the coupled assay with tiglyl-CoA.
2)We identified a large T2 gene deletion including exons 2-4 caused by unequal homologous recombination. This is the first mutation caused by this mechanism in T2 gene.
3.Liver specific suppression of human SCOT gene expression.
We analyzed status of methylation in CpG islands around SCOT gene promotor regions in HeLa cells and hepatoblastoma cell lines and murine hepatic tissues and cardiac muscle. Unexpectedly, the CpG islands around SCOT promotor regions were hypomethylated in even hepatoblastoma cell lines and murine hepatic tissues where SCOT gene was almost completely suppressed.
4.Tertiary structure of human thiolase family.
We determined a crystal tertiary structure of cytosolic acetoacetyl-CoA thiolase and found several new findings in enzyme mechamism using the structure.

Research Products

• [Journal Article] Regulation of the Atm promoter in vivo. (2006)
  • Author(s): Gueven N
  • Journal Title: Genes Chromosomes Cancer. 45 Pages: 61-71
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Regulation of the Atm promoter in vivo. (2006)
  • Author(s): Gueven N, Fukao T, Luff J, Paterson C, Kay G, Kondo N, Lavin MF.
  • Journal Title: Genes Chromosomes Cancer. 45(1) Pages: 61-71
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Regulation of the Atm promoter in vivo. (2006)
  • Author(s): Gueven N, Fukao T, Luff J, Paterson C, Kay G, Kondo N, Lavin MF.
  • Journal Title: Genes Chromosomes Cancer 45 Pages: 61-71
• [Journal Article] High resolution crystal structures of human cytosolic thiolase (CT). A comparison of the active sites of human CT, bacterial thiolase, and bacterial KAS I (2005)
  • Author(s): Kursula P
  • Journal Title: J Mol Biol 347 Pages: 189-201
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Two novel mutations in mitochondrial acetoacetyl-CoA thiolase deficiency (2005)
  • Author(s): Mrazova L
  • Journal Title: J Inherit Metab Dis 28 Pages: 235-236
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Questionnaire-based study on the relationship between pet-keeping and allergic diseases in young children in Japan. (2005)
  • Author(s): Fukao T
  • Journal Title: Allergol International 54 Pages: 521-526
  • NAID: 10016880688
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of T-cell clones specific to ovomucoid from patients with egg-white allergy. (2005)
  • Author(s): Kondo M
  • Journal Title: J Investig Allergol Clin Immunol 15 Pages: 107-111
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A common variable immunodeficient patient who developed acute disseminated encephalomyelitis followed by the Lennox-Gastaut syndrome. (2005)
  • Author(s): Kondo M
  • Journal Title: Pediatr Allergy Immunol. 16 Pages: 357-360
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] IL-12B promotor polymorphism associated with asthma and IL-12B transcriptional activity. (2005)
  • Author(s): Tatebayashi K.
  • Journal Title: Allergol International 54 Pages: 345-349
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A novel single-nucleotide substitution, Glu 4 Lys, in the leukotriene C4 synthase gene associated with allergic diseases. (2005)
  • Author(s): Yoshikawa K
  • Journal Title: Int J Mol Med 16 Pages: 827-831
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Leaky phenotype of X-linked agammaglobulinaemia in a Japanese family. (2005)
  • Author(s): Kaneko H
  • Journal Title: Clin Exp Immunol. 140 Pages: 520-523
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Mutational and structural analysis of Japanese patients with mucopolysaccharidosis type II. (2005)
  • Author(s): Kato T
  • Journal Title: J Hum Genet 50 Pages: 395-402
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] High resolution crystal structures of human cytosolic thiolase (CT). A comparison of the active sites of human CT, bacterial thiolase, and bacterial KAS I. (2005)
  • Author(s): Kursula P, Sikkila H, Fukao T, Kondo N, Wierenga RK
  • Journal Title: J Mol Biol. 347 Pages: 189-201
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of T-cell clones specific to ovomucoid from patients with egg-white allergy. (2005)
  • Author(s): Kondo M, Suzuki K, Inoue R, Sakaguchi H, Matsukuma E, Kato Z, Kaneko H, Fukao T, Kondo N
  • Journal Title: J Investig Allergol Clin Immunol. 15 Pages: 107-111
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A common variable immunodeficient patient who developed acute disseminated encephalomyelitis followed by the Lennox-Gastaut syndrome. (2005)
  • Author(s): Kondo M, Fukao T, Teramoto T, Takahashi Y, Okamoto H, Kondo N
  • Journal Title: Pediatr Allergy Immunol. 16 Pages: 357-360
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Leaky phenotype of X-linked agammaglobulinaemia in a Japanese family. (2005)
  • Author(s): Kaneko H, Kawamoto N, Asano T, Mabuchi Y, Horikoshi H, Teramoto T, Matsui E, Kondo M, Fukao T, Kasahara K, Kondo N.
  • Journal Title: Clin Exp Immunol. 140 Pages: 520-523
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two novel mutations in mitochondrial acetoacetyl-CoA thiolase deficiency (2005)
  • Author(s): Mrazova L, Fukao T, Halovd K, Gregova E, Kohut V, Pribyl D, Chrastina P, Kondo N, Pospisilova E.
  • Journal Title: J Inherit Metab Dis. 28 Pages: 235-236
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] IL-12B promotor polymorphism associated with asthma and IL-12B transcriptional activity. (2005)
  • Author(s): Tatebayashi K, Matsui E, Kaneko H, Fukao T, Kasahara K, Kondo N.
  • Journal Title: Allergol Int 54 Pages: 345-349
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Mutational and structural analysis of Japanese patients with mucopolysaccharidosis type II. (2005)
  • Author(s): Kato T, Kato Z, Kuratsubo I, Tanaka N, Ishigami T, Kajihara J, Sukegawa-Hayasaka K, Orii K, Isogai K, Fukao T, Shimozawa N, Orii T, Kondo N, Suzuki Y.
  • Journal Title: J Hum Genet 50 Pages: 395-402
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A novel single-nucleotide substitution, Glu 4 Lys, in the leukotriene C4 synthase gene associated with allergic diseases. (2005)
  • Author(s): Yoshikawa K, Matsui E, Kaneko H, Fukao T, Inoue R, Teramoto T, Shinoda S, Fukutomi O, Aoki M, Kasahara K, Kondo N.
  • Journal Title: Int J Mol Med. 16 Pages: 827-831
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Questionnaire-based study on the relationship between pet-keeping and allergic diseases in young children in Japan. (2005)
  • Author(s): Fukao T, Fukutomi O, Hirayama K, Teramoto T, Kaneko H, Kondo M, Matsui E, Kondo N.
  • Journal Title: Allergol International 54 Pages: 521-526
  • NAID: 10016880688
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] High resolution crystal structures of human cytosolic thiolase (CT). A comparison of the active sites of human CT, bacterial thiolase, and bacterial KASI (2005)
  • Author(s): Kursula P, Sikkila H, Fukao T, Kondo N, Wierenga RK
  • Journal Title: J Mol Biol 347 Pages: 189-201
• [Journal Article] Two novel mutations in mitochondrial acetoacetyl-CoA thiolase deficiency (2005)
  • Author(s): Mrazova L, Fukao T, Halovd K, Gregova E, Kohut V, Pribyl D, Chrastina P, Kondo N, Pospisilova E
  • Journal Title: J Inherit Metab Dis 28 Pages: 235-236
• [Journal Article] Characterization of T-cell clones specific to ovomucoid from patients with egg-white allergy. (2005)
  • Author(s): Kondo M, Suzuki K, Inoue R, Sakaguchi H, Matsukuma E, Kato Z, Kaneko H, Fukao T, Kondo N
  • Journal Title: J Investig Allergol Clin Immunol 15 Pages: 107-111
• [Journal Article] IL-12B promotor polymorphism associated with asthma and IL-12B transcriptional activity. (2005)
  • Author(s): Tatebayashi K, Matsui E, Kaneko H, Fukao T, Kasahara K, Kondo N.
  • Journal Title: Allergol International 54 Pages: 345-349
• [Journal Article] A novel single-nucleotide substitution, Glu 4 Lys, in the leukotriene C4 synthase gene associated with allergic diseases. (2005)
  • Author(s): Yoshikawa K, Matsui E, Kaneko H, Fukao T, Inoue R, Teramoto T, Shinoda S, Fukutomi O, Aoki M, Kasahara K. Kbndo N.
  • Journal Title: Int J Mol Med 16 Pages: 827-831
• [Journal Article] Leaky phenotype of X-linked agammaglobulinaemia in a Japanese family. (2005)
  • Author(s): Kaneko H, Kawamoto N, Asano T, Mabuchi Y, Horikoshi H, Teramoto T, Matsui E, Kondo M, Fukao T, Kasahara K, Kondo N
  • Journal Title: Clin Exp Immunol. 140 Pages: 520-523
• [Journal Article] Mutational and structural analysis of Japanese patients with mucopolysaccharidosis type II. (2005)
  • Author(s): Kato T, Kato Z, Kuratsubo I, Tanaka N, Ishigami T, Kajihar J, Sukegawa-Hayasaka K, Orii K, Isogai K, Fukao T, Shimozawa N, Orii T, Kondo N, Suzuki Y.
  • Journal Title: J Hum Genet 50 Pages: 395-402
• [Journal Article] Patients homozygous for the T435N mutation of succinyl-CoA : 3-ketoacid CoA transferase (SCOT) do not show permanent ketosis. (2004)
  • Author(s): Fukao T
  • Journal Title: Pediatr Res 56 Pages: 858-863
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Succinyl-CoA : 3-keto acid transferase (SCOT) deficiency in a new patient homozygous for an R217X mutation. (2004)
  • Author(s): Longo N
  • Journal Title: J Inherit Metab Dis 27 Pages: 691-692
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The mitochondrial acetoacetyl-CoA thiolase (T2) deficiency : T2-deficient patients with メ mild モ mutation(s) were previously misinterpreted as normal by the coupled assay with tiglyl-CoA (2004)
  • Author(s): Zhang G-X
  • Journal Title: Pediatr Res 56 Pages: 60-64
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Relatively common mutations of the Bloom syndrome gene in the Japanese population (2004)
  • Author(s): Kaneko H
  • Journal Title: Int J Mol Med 14 Pages: 439-442
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of somatic and germ-line mosaicism for a keratin 5 mutation in epidermolysis bullosa simplex in a family of which the proband was previously regarded as a sporadic case. (2004)
  • Author(s): Nagao-Watanabe M
  • Journal Title: Clin Genet 66 Pages: 236-238
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] RNA editing of interleukin-12 receptor beta2, 2451 C-to-U (Ala 604 Val) conversion, associated with atopy. (2004)
  • Author(s): Kondo N
  • Journal Title: Clin Exp Allergy 34 Pages: 363-368
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Molecular analysis of B-cell differentiation in selective or partial IgA deficiency. (2004)
  • Author(s): Asano T
  • Journal Title: Clin Exp Immunol 136 Pages: 284-290
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Disruption of the BLM gene in ATM-null DT40 cells does not exacerbate either phenotype (2004)
  • Author(s): Fukao T
  • Journal Title: Oncogene 23 Pages: 1498-1506
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Pathways and control of ketone body metabolism : on the fringe of lipid biochemistry. (2004)
  • Author(s): Fukao T
  • Journal Title: Prostaglandins, Leukotrienes and Essential Fatty Acid 70 Pages: 77-85
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Patients homozygous for the T435N mutation of succinyl-CoA:3-ketoacid CoA transferase (SCOT) do not show permanent ketosis. (2004)
  • Author(s): Fukao T, Shintaku H, Kusubae R, Zhang X-Q, Nakamura K, Kondo M, Kondo N.
  • Journal Title: Pediatr Res 56 Pages: 858-863
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The mitochondrial acetoacetyl-CoA thiolase (T2) deficiency : T2-deficient patients with メ mild モ mutation(s) were previously misinterpreted as normal by the coupled assay with tiglyl-CoA. (2004)
  • Author(s): Zhang G-X, Fukao T, Rolland M-O, Zabot M-T, Renom G, Touma E, Kondo M, Matsuo N, Kondo N
  • Journal Title: Pediatr Res 56 Pages: 60-64
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Disruption of the BLM gene in ATM-null DT40 cells does not exacerbate either phenotype. (2004)
  • Author(s): Fukao T, Chen P, Ren J, Kaneko H, Zhang GX, Kondo M, Yamamoto K, Furuichi Y, Takeda S, Kondo N, Lavin MF
  • Journal Title: Oncogene 23 Pages: 1498-1506
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular analysis of B-cell differentiation in selective or partial IgA deficiency. (2004)
  • Author(s): Asano T, Kaneko H, Terada T, Kasahara Y, Fukao T, Kasahara K, Kondo N
  • Journal Title: Clin Exp Immunol 136 Pages: 284-290
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] RNA editing of interleukin-12 receptor beta2, 2451 C-to-U (Ala 604 Val) conversion, associated with atopy. (2004)
  • Author(s): Kondo N, Matsui E, Kaneko H, Aoki M, Kato Z, Fukao T, Kasahara K, Morimoto N
  • Journal Title: Clin Exp Allergy 34(3) Pages: 363-368
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of somatic and germ-line mosaicism for a keratin 5 mutation in epidermolysis bullosa simplex in a family of which the proband was previously regarded as a sporadic case. (2004)
  • Author(s): Nagao-Watanabe M, Fukao T, Matsui E, Kaneko H, Inoue R, Kawamoto N, Kasahara K, Nagai M, Ichiki Y, Kitajima Y, Kondo N
  • Journal Title: Clin Genet 66 Pages: 236-238
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Relatively common mutations of the Bloom syndrome gene in the Japanese population. (2004)
  • Author(s): Kaneko H, Isogai K, Fukao T, Matsui E, Kasahara K, Yachie A, Seki H, Koizumi S, Arai M, Utunomiya J, Miki Y, Kondo N
  • Journal Title: Int J Mol Med 14 Pages: 439-442
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Succinyl-CoA:3-keto acid transferase (SCOT) deficiency in a new patient homozygous for an R217X mutation. (2004)
  • Author(s): Longo N, Fukao T, Singh R, Pasquali M, Barrios RG, Kondo N, Gibson KM
  • Journal Title: J Inherit Metab Dis 27 Pages: 691-692
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The mitochondrial acetoacetyl- CoA thiolase (T2) deficiency : T2-deficient patients with "mild" mutation(s) were previously misinterpreted as normal by the coupled assay with tiglyl-CoA (2004)
  • Author(s): Zhang G-X, Fukao T, et al.
  • Journal Title: Pediatr Res 56 Pages: 60-64
• [Journal Article] Disruption of the BLM gene in ATM-null DT40 cells does not exacerbate either phenotyne. (2004)
  • Author(s): Fukao T, et al.
  • Journal Title: Oncogene 23 Pages: 1498-1506
• [Journal Article] Pathways and control of ketone body metabolism : on the fringe of lipid biochemisitry. (2004)
  • Author(s): Fukao T, Lopaschuk GD, Mitchell GA
  • Journal Title: Prostaglandins, Leukotrienes and Essential Fatty Acid 70 Pages: 243-251
• [Journal Article] Patients homozygous for the T435N mutation of succinyl-CoA:3-keto acid CoA transferase (SCOT) do not show permanent ketosis (2004)
  • Author(s): Fukao T, et al.
  • Journal Title: Pediatr Res 56 Pages: 858-863
• [Journal Article] Identification of somatic and germ-line mosaicism for a keratin 5 mutation in epidermolysis bullosa simplex in a family of which the proband was previously regarded as a sporadic case. (2004)
  • Author(s): Nagao-Watanabe M, Fukao T, et al.
  • Journal Title: Clin Genet 66 Pages: 236-238
• [Journal Article] Succinyl-CoA:3-keto acid transferase (SCOT) deficiency in a new patient homozygous for an R217X mutation. (2004)
  • Author(s): Longo N, Fukao T, et al.
  • Journal Title: J Inherit Metab Dis 27 Pages: 691-692
• [Journal Article] Aberrant peroxisome morphology in peroxisomal beta-oxidation enzyme deficiencies.
  • Author(s): Funato M, Shimozawa N, Nagase T, Takemoto Y, Suzuki Y, Imamura Y, Matsumoto T, Tsukamoto T, Kojidani T, Osumi T, Fukao T, Kondo N.
  • Journal Title: Brain Dev. (In press)
  • NAID: 10020330709
  • Description: 「研究成果報告書概要(欧文)」より