• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Ubiquitination of Niemann-Pick Cl protein

Research Project

Project/Area Number 18590295
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Pathological medical chemistry
Research InstitutionTottori University

Principal Investigator

NINOMIYA Haruaki  Tottori University, Faculty of Medicine, Professor (80212124)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥4,010,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsNiemann-Pick / cholesterol / ubiquitin / roteasome
Research Abstract

We and others reported reduced protein levels of NPC1 in fibroblasts from patients with NPC1 mutations. Importantly, cells from patients with juvenile/adult forms retained relatively high levels of the protein, suggesting that the NPC1 protein level may be one of the factors that determine the disease severity. We found that a proteasome inhibitor MG132 caused an accumulation of ubiquitinated NPC1, suggesting proteasomal degradation of NPC1s. Pulse-chase analysis in COS cells revealed that an I1061T mutation decreased the half-life time of expressed NPC1 protein. Wild-type as well as loss-of-function mutant NPC1 proteins associated with molecular chaperones HSP70, HSP90 and calnexin. Accordingly, overexpression of HSP70 in human fibroblasts with I1061T homozygous mutations by adeno-HSP70 or treatment with geranylgeranylacetone increased the level of the mutant protein. In COS cells, co-expression of an E3 ligase CHIP (carboxyl terminus of HSP70-interacting protein) enhanced MG132-induced accumulation of ubiquitinated NPC1. MALDI-TOF mass spectrometry has revealed three lysine residues on the cytosolic side, K318, K792 and K1180, as potential acceptors of ubiquitin. Substitution of the three lysine residues with alanine yielded a mutant protein with a steady-state level approximately 10 times higher than the wild-type protein. These findings indicate that NPC1 undergoes proteasomal degradation and that its biosynthesis is an inefficient process: 90% of the wild-type protein is degraded away. These findings also suggest the possibility to restore endosomal cholesterol flow by stabilization of the mutant NPC1 proteins.

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (19 results)

All 2007 2006 Other

All Journal Article (14 results) (of which Peer Reviewed: 5 results) Presentation (3 results) Remarks (2 results)

  • [Journal Article] Endosomal accumulation of Toll-like receptor 4 causes constitutive secretion of cytokines and activation of signal transducers and activators of transcription in Niemann-Pick disease type C (NPC) fibroblasts: a potential basis for glial cell activation in the NPC brain.2007

    • Author(s)
      Suzuki, et. al.
    • Journal Title

      J. Neurosci. 27

      Pages: 1879-1891

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Amyloid beta inhibits ectodomain shedding of N-cadherin via down-regulation of cell-surface NMDA receptor.2007

    • Author(s)
      Uemura, et. al.
    • Journal Title

      Neuroscience 145

      Pages: 5-10

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Presenilin 1 is involved in the maturation of beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1).2007

    • Author(s)
      Kuzuya, et. al.
    • Journal Title

      J. Neurosci. Res. 85

      Pages: 153-165

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Endosomal accumulation of Toll-like receptor 4 causes constitutive secretion of cytokines and activation of signal transducers and activators of transcription in Niemann-Pick disease type C(NPC) fibroblasts: a potential basis for glial cell activation in the NPC brain2007

    • Author(s)
      Suzuki, et. al.
    • Journal Title

      J. Neurosci 27

      Pages: 1879-1891

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Amyloid beta inhibits ectodomain shedding of N-cadherin via down-regulation of cell-surface NMDAreceptor2007

    • Author(s)
      Uemura, et. al.
    • Journal Title

      Neuroscience 145

      Pages: 5-10

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Presenilin 1 is involved in the maturation of beta-site amyloid precursor protein-cleaving enzyme 1(BACE1)2007

    • Author(s)
      Kuzuya, et. al.
    • Journal Title

      J. Neurosci. Res 85

      Pages: 153-165

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Endosomal accumulation of Toll-like receptor 4 causes constitutive secretion of cytokines and activation of signal transducers and activators of transcription in Niemann-Pick disease type C(NPC)fibroblasts: a potential basis for glial cell activationin the NPC brain.2007

    • Author(s)
      Suzuki, et. al.
    • Journal Title

      J. Neurosci. 27

      Pages: 1879-1891

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Presenilin 1 is involved in the maturation of beta-site amyloid precursor protein-cleaving enzyme 1(BACE1).2007

    • Author(s)
      Kuzuya, et. al.
    • Journal Title

      J. Neurosci. Res. 85

      Pages: 153-165

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Inhibition of inward rectifier K^+ currents by angiotensin II in rat atrial myocytes : lack of effects in cells from spontaneous hypertensive rats.2006

    • Author(s)
      Sonoyma K, et al.
    • Journal Title

      Hypertension Res. 29

      Pages: 923-934

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Delayed onset of beating and decreased expression of T-type Ca^2+ channel in mouse ES cell-derived cardiocytes carrying human chromosome 21.2006

    • Author(s)
      Mizuta E, et al.
    • Journal Title

      Biochem. Biophys. Res. Commun. 351

      Pages: 126-132

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Cholesterol depletion facilitates ubiquitylation of NPC1 and its association with SKD1/Vps4.2006

    • Author(s)
      Ohsaki Y, et al.
    • Journal Title

      J. Cell Sci. 199

      Pages: 2643-2653

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Activity-dependent regulation of beta-catenin via epsilon-cleavage of N-cadherin.2006

    • Author(s)
      Uemura K, et al.
    • Journal Title

      Biochem. Biophys. Res. Commun. 345

      Pages: 951-958

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Characterization of sequential N-cadherin cleavage by ADAM10 and PS1.2006

    • Author(s)
      Uemura K, et al.
    • Journal Title

      Neurosci. Lett. 402

      Pages: 278-283

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Niemann-Pick type C disease : Novel NPC1 mutations and characterization of the concomitant acid sphingomyelinase deficiency.2006

    • Author(s)
      Tamura H, et al.
    • Journal Title

      Mol. Genet. Metab. 87

      Pages: 113-121

    • Related Report
      2006 Annual Research Report
  • [Presentation] Proteasomal degradation of NPCl.2007

    • Author(s)
      Ninomiya H.
    • Organizer
      Annual Scientific Conference on NPC
    • Place of Presentation
      Tucson, U.S.A.
    • Year and Date
      2007-06-07
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Proteasomal degradation of NPC1.2007

    • Author(s)
      Ninomiya H.
    • Organizer
      Annual Scientific Conference on NPC
    • Place of Presentation
      Tucson, U. S. A.
    • Year and Date
      2007-06-07
    • Related Report
      2007 Annual Research Report
  • [Presentation] Proteasomal degradation of NPC12007

    • Author(s)
      Ninomiya, H
    • Organizer
      Annual Scientific Conference on NPC
    • Place of Presentation
      Tucson, U.S.A
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Remarks] 「研究成果報告書概要(和文)」より

    • URL

      http://www.asahi-net.or.jp/~ev6h-nnmy/

    • Related Report
      2007 Final Research Report Summary
  • [Remarks]

    • URL

      http://www.asahi-net.or.jp/~ev6h-nnmy/

    • Related Report
      2007 Annual Research Report

URL: 

Published: 2006-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi