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Vol. 18, No. 4, 2010
Issue release date: March 2011
Section title: Original Paper
Free Access
Neurosignals 2010;18:203–209
(DOI:10.1159/000322206)

Association of the Small GTPase Rheb with the NMDA Receptor Subunit NR3A

Sucher N.J.a–c · Yu E.d · Chan S.F.d · Miri M.b · Lee B.J.b · Xiao B.e · Worley P.F.e · Jensen F.E.b, c, f
aCentre for Complementary Medicine Research, University of Western Sydney, Penrith South DC, N.S.W., Australia; bDepartment of Neurology, Division of Neuroscience, Children’s Hospital, and cDepartment of Neurology, Harvard Medical School, Boston, Mass., dSanford-Burnham Medical Research Institute, La Jolla, Calif. eDepartment of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Md., and fProgram in Neuroscience, Harvard Medical School, Boston, Mass., USA
email Corresponding Author

Abstract

The NMDAR subunit NR3A is most highly expressed during the second postnatal week, when synaptogenesis reaches peak levels. Genetic ablation or overexpression of the NR3A subunit negatively interferes with the maturation of cortical synapses and leads to changes in the shape and number of dendritic spines, the density of which is increased in NR3A knock-out mice and decreased in NR3A-overexpressing transgenic mice. Alterations in spine density have been linked to dysregulation of mTOR signaling and synaptic protein translation. Using a yeast two-hybrid system, we identified the mTOR-activating GTPase Rheb as an interacting protein of the NMDAR subunit NR3A. We confirmed the interaction in mammalian cells by expressing recombinant Rheb and NR3A and showed that Rheb and NR3A could be co-immunoprecipitated from synaptic plasma membranes from the developing rat brain. These data suggest that NR3A sequesters synaptic Rheb and might thus function as a break of the mTOR-dependent synaptic translation of protein.

© 2010 S. Karger AG, Basel


  

Key Words

  • NMDA receptor
  • Rheb
  • Synapse
  • Plasticity
  • Translation
  • mTOR

References

  1. Feldman DE: Synaptic mechanisms for plasticity in neocortex. Annu Rev Neurosci 2009;32:33–55.
  2. Malenka RC, Bear MF: LTP and LTD: An embarrassment of riches. Neuron 2004;44:5–21.
  3. Carlisle HJ, Kennedy MB: Spine architecture and synaptic plasticity. Trends Neurosci 2005;28:182–187.
  4. Hoeffer CA, Klann E: mTOR signaling: at the crossroads of plasticity, memory and disease. Trends Neurosci 2010;33:67–75.
  5. Cavara NA, Hollmann M: Shuffling the deck anew: how NR3A tweaks NMDA receptor function. Mol Neurobiol 2008;38:16–26.
  6. Cull-Candy S, Brickley S, Farrant M: NMDA receptor subunits: diversity, development and disease. Curr Opin Neurobiol 2001;11:327–335.
  7. Wong H-K, Liu X-B, Matos MF, Chan SF, Pérez-Otaño I, Boysen M, Cui J, Nakanishi N, Trimmer JS, Jones EG, Lipton SA, Sucher NJ: Temporal and regional expression of NMDA receptor subunit NR3A in the mammalian brain. J Comp Neurol 2002;450:303–317.
  8. Zhou C, Jensen F, Sucher N: Altered development of glutamatergic synapses in layer V pyramidal neurons in NR3A knockout mice. Mol Cell Neurosci 2009;42:419–426.
  9. Das S, Sasaki YF, Rothe T, Premkumar LS, Takasu M, Crandall JE, Dikkes P, Conner DA, Rayudu PV, Cheung W, Chen HS, Lipton SA, Nakanishi N: Increased NMDA current and spine density in mice lacking the NMDA receptor subunit NR3A. Nature 1998;393:377–381.
  10. Roberts AC, Díez-García J, Rodriguiz RM, López IP, Luján R, Martínez-Turrillas R, Picó E, Henson MA, Bernardo DR, Jarrett TM, Clendeninn DJ, López-Mascaraque L, Feng G, Lo DC, Wesseling JF, Wetsel WC, Philpot BD, Pérez-Otaño I: Downregulation of NR3A-containing NMDARs is required for synapse maturation and memory consolidation. Neuron 2009;63:342–356.
  11. Bassell GJ, Warren ST: Fragile X syndrome: loss of local mRNA regulation alters synaptic development and function. Neuron 2008;60:201–214.
  12. Bear MF, Dolen G, Osterweil E, Nagarajan N: Fragile X: Translation in action. Neuropsychopharmacology 2008;33:84–87.
  13. Narayanan U, Nalavadi V, Nakamoto M, Pallas DC, Ceman S, Bassell GJ, Warren ST: FMRP phosphorylation reveals an immediate-early signaling pathway triggered by group I mGluR and mediated by PP2A. J Neurosci 2007;27:14349–14357.
  14. Narayanan U, Nalavadi V, Nakamoto M, Thomas G, Ceman S, Bassell GJ, Warren ST: S6k1 phosphorylates and regulates fragile X mental retardation protein (FMRP) with the neuronal protein synthesis-dependent mammalian target of rapamycin (mTOR) signaling cascade. J Biol Chem 2008;283:18478–18482.
  15. Arsham AM, Neufeld TP: Thinking globally and acting locally with TOR. Curr Opin Cell Biol 2006;18:589–597.
  16. Chan SF, Sucher NJ: An NMDA receptor signaling complex with protein phosphatase 2a. J Neurosci 2001;21:7985–7992.
  17. Yamagata K, Sanders LK, Kaufmann WE, Yee W, Barnes CA, Nathans D, Worley PF: Rheb, a growth factor- and synaptic activity-regulated gene, encodes a novel ras-related protein. J Biol Chem 1994;269:16333–16339.
  18. Aspuria P-J, Tamanoi F: The Rheb family of GTP-binding proteins. Cell Signal 2004;16:1105–1112.
  19. Ma OK, Sucher NJ: Molecular interaction of NMDA receptor subunit NR3A with protein phosphatase 2A. Neuroreport 2004;15:1447–1450.
  20. Slabinski L, Jaroszewski L, Rodrigues AP, Rychlewski L, Wilson IA, Lesley SA, Godzik A: The challenge of protein structure determination – lessons from structural genomics. Protein Sci 2007;16:2472–2482.
  21. Slabinski L, Jaroszewski L, Rychlewski L, Wilson IA, Lesley SA, Godzik A: Xtalpred: a web server for prediction of protein crystallizability. Bioinformatics 2007;23:3403–3405.
  22. Igarashi Y, Eroshkin A, Gramatikova S, Gramatikoff K, Zhang Y, Smith JW, Osterman AL, Godzik A: Cutdb: a proteolytic event database. Nucleic Acids Res 2007;35:D546–D549.
  23. Li N, Lee B, Liu RJ, Banasr M, Dwyer JM, Iwata M, Li XY, Aghajanian G, Duman RS: mTOR-dependent synapse formation underlies the rapid antidepressant effects of NMDA antagonists. Science 2010;329:959–964.
  24. Tang SJ, Schuman EM: Protein synthesis in the dendrite. Philos Trans R Soc Lond B Biol Sci 2002;357:521–529.
  25. Sharma A, Hoeffer CA, Takayasu Y, Miyawaki T, McBride SM, Klann E, Zukin RS: Dysregulation of mTOR signaling in fragile X syndrome. J Neurosci 2010;30:694–702.
  26. Weatherill DB, Dyer J, Sossin WS: Ribosomal protein S6 kinase is a critical downstream effector of the target of rapamycin complex 1 for long-term facilitation in aplysia. J Biol Chem 2010;285:12255–12267 .
  27. Lee MN, Ha SH, Kim J, Koh A, Lee CS, Kim JH, Jeon H, Kim DH, Suh PG, Ryu SH: Glycolytic flux signals to mTOR through glyceraldehyde-3-phosphate dehydrogenase-mediated regulation of Rheb. Mol Cell Biol 2009;29:3991–4001.

  

Author Contacts

Prof. Nikolaus J. Sucher
The Centre for Complementary Medicine Research
University of Western Sydney, Locked Bag 1797
Penrith, NSW 2751 (Australia)
Tel. +61 2 4620 3345, Fax +61 2 4620 3017, E-Mail n.sucher@uws.edu.au

  

Article Information

Received: July 27, 2010
Accepted after revision: October 20, 2010
Published online: December 7, 2010
Number of Print Pages : 7
Number of Figures : 3, Number of Tables : 0, Number of References : 27

  

Publication Details

Neurosignals

Vol. 18, No. 4, Year 2010 (Cover Date: March 2011)

Journal Editor: Ip N.Y. (Hong Kong)
ISSN: 1424-862X (Print), eISSN: 1424-8638 (Online)

For additional information: http://www.karger.com/NSG


Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Abstract

The NMDAR subunit NR3A is most highly expressed during the second postnatal week, when synaptogenesis reaches peak levels. Genetic ablation or overexpression of the NR3A subunit negatively interferes with the maturation of cortical synapses and leads to changes in the shape and number of dendritic spines, the density of which is increased in NR3A knock-out mice and decreased in NR3A-overexpressing transgenic mice. Alterations in spine density have been linked to dysregulation of mTOR signaling and synaptic protein translation. Using a yeast two-hybrid system, we identified the mTOR-activating GTPase Rheb as an interacting protein of the NMDAR subunit NR3A. We confirmed the interaction in mammalian cells by expressing recombinant Rheb and NR3A and showed that Rheb and NR3A could be co-immunoprecipitated from synaptic plasma membranes from the developing rat brain. These data suggest that NR3A sequesters synaptic Rheb and might thus function as a break of the mTOR-dependent synaptic translation of protein.

© 2010 S. Karger AG, Basel


  

Author Contacts

Prof. Nikolaus J. Sucher
The Centre for Complementary Medicine Research
University of Western Sydney, Locked Bag 1797
Penrith, NSW 2751 (Australia)
Tel. +61 2 4620 3345, Fax +61 2 4620 3017, E-Mail n.sucher@uws.edu.au

  

Article Information

Received: July 27, 2010
Accepted after revision: October 20, 2010
Published online: December 7, 2010
Number of Print Pages : 7
Number of Figures : 3, Number of Tables : 0, Number of References : 27

  

Publication Details

Neurosignals

Vol. 18, No. 4, Year 2010 (Cover Date: March 2011)

Journal Editor: Ip N.Y. (Hong Kong)
ISSN: 1424-862X (Print), eISSN: 1424-8638 (Online)

For additional information: http://www.karger.com/NSG


Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: 7/27/2010
Accepted: 10/20/2010
Published online: 12/7/2010
Issue release date: March 2011

Number of Print Pages: 7
Number of Figures: 3
Number of Tables: 0

ISSN: 1424-862X (Print)
eISSN: 1424-8638 (Online)

For additional information: http://www.karger.com/NSG


Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Feldman DE: Synaptic mechanisms for plasticity in neocortex. Annu Rev Neurosci 2009;32:33–55.
  2. Malenka RC, Bear MF: LTP and LTD: An embarrassment of riches. Neuron 2004;44:5–21.
  3. Carlisle HJ, Kennedy MB: Spine architecture and synaptic plasticity. Trends Neurosci 2005;28:182–187.
  4. Hoeffer CA, Klann E: mTOR signaling: at the crossroads of plasticity, memory and disease. Trends Neurosci 2010;33:67–75.
  5. Cavara NA, Hollmann M: Shuffling the deck anew: how NR3A tweaks NMDA receptor function. Mol Neurobiol 2008;38:16–26.
  6. Cull-Candy S, Brickley S, Farrant M: NMDA receptor subunits: diversity, development and disease. Curr Opin Neurobiol 2001;11:327–335.
  7. Wong H-K, Liu X-B, Matos MF, Chan SF, Pérez-Otaño I, Boysen M, Cui J, Nakanishi N, Trimmer JS, Jones EG, Lipton SA, Sucher NJ: Temporal and regional expression of NMDA receptor subunit NR3A in the mammalian brain. J Comp Neurol 2002;450:303–317.
  8. Zhou C, Jensen F, Sucher N: Altered development of glutamatergic synapses in layer V pyramidal neurons in NR3A knockout mice. Mol Cell Neurosci 2009;42:419–426.
  9. Das S, Sasaki YF, Rothe T, Premkumar LS, Takasu M, Crandall JE, Dikkes P, Conner DA, Rayudu PV, Cheung W, Chen HS, Lipton SA, Nakanishi N: Increased NMDA current and spine density in mice lacking the NMDA receptor subunit NR3A. Nature 1998;393:377–381.
  10. Roberts AC, Díez-García J, Rodriguiz RM, López IP, Luján R, Martínez-Turrillas R, Picó E, Henson MA, Bernardo DR, Jarrett TM, Clendeninn DJ, López-Mascaraque L, Feng G, Lo DC, Wesseling JF, Wetsel WC, Philpot BD, Pérez-Otaño I: Downregulation of NR3A-containing NMDARs is required for synapse maturation and memory consolidation. Neuron 2009;63:342–356.
  11. Bassell GJ, Warren ST: Fragile X syndrome: loss of local mRNA regulation alters synaptic development and function. Neuron 2008;60:201–214.
  12. Bear MF, Dolen G, Osterweil E, Nagarajan N: Fragile X: Translation in action. Neuropsychopharmacology 2008;33:84–87.
  13. Narayanan U, Nalavadi V, Nakamoto M, Pallas DC, Ceman S, Bassell GJ, Warren ST: FMRP phosphorylation reveals an immediate-early signaling pathway triggered by group I mGluR and mediated by PP2A. J Neurosci 2007;27:14349–14357.
  14. Narayanan U, Nalavadi V, Nakamoto M, Thomas G, Ceman S, Bassell GJ, Warren ST: S6k1 phosphorylates and regulates fragile X mental retardation protein (FMRP) with the neuronal protein synthesis-dependent mammalian target of rapamycin (mTOR) signaling cascade. J Biol Chem 2008;283:18478–18482.
  15. Arsham AM, Neufeld TP: Thinking globally and acting locally with TOR. Curr Opin Cell Biol 2006;18:589–597.
  16. Chan SF, Sucher NJ: An NMDA receptor signaling complex with protein phosphatase 2a. J Neurosci 2001;21:7985–7992.
  17. Yamagata K, Sanders LK, Kaufmann WE, Yee W, Barnes CA, Nathans D, Worley PF: Rheb, a growth factor- and synaptic activity-regulated gene, encodes a novel ras-related protein. J Biol Chem 1994;269:16333–16339.
  18. Aspuria P-J, Tamanoi F: The Rheb family of GTP-binding proteins. Cell Signal 2004;16:1105–1112.
  19. Ma OK, Sucher NJ: Molecular interaction of NMDA receptor subunit NR3A with protein phosphatase 2A. Neuroreport 2004;15:1447–1450.
  20. Slabinski L, Jaroszewski L, Rodrigues AP, Rychlewski L, Wilson IA, Lesley SA, Godzik A: The challenge of protein structure determination – lessons from structural genomics. Protein Sci 2007;16:2472–2482.
  21. Slabinski L, Jaroszewski L, Rychlewski L, Wilson IA, Lesley SA, Godzik A: Xtalpred: a web server for prediction of protein crystallizability. Bioinformatics 2007;23:3403–3405.
  22. Igarashi Y, Eroshkin A, Gramatikova S, Gramatikoff K, Zhang Y, Smith JW, Osterman AL, Godzik A: Cutdb: a proteolytic event database. Nucleic Acids Res 2007;35:D546–D549.
  23. Li N, Lee B, Liu RJ, Banasr M, Dwyer JM, Iwata M, Li XY, Aghajanian G, Duman RS: mTOR-dependent synapse formation underlies the rapid antidepressant effects of NMDA antagonists. Science 2010;329:959–964.
  24. Tang SJ, Schuman EM: Protein synthesis in the dendrite. Philos Trans R Soc Lond B Biol Sci 2002;357:521–529.
  25. Sharma A, Hoeffer CA, Takayasu Y, Miyawaki T, McBride SM, Klann E, Zukin RS: Dysregulation of mTOR signaling in fragile X syndrome. J Neurosci 2010;30:694–702.
  26. Weatherill DB, Dyer J, Sossin WS: Ribosomal protein S6 kinase is a critical downstream effector of the target of rapamycin complex 1 for long-term facilitation in aplysia. J Biol Chem 2010;285:12255–12267 .
  27. Lee MN, Ha SH, Kim J, Koh A, Lee CS, Kim JH, Jeon H, Kim DH, Suh PG, Ryu SH: Glycolytic flux signals to mTOR through glyceraldehyde-3-phosphate dehydrogenase-mediated regulation of Rheb. Mol Cell Biol 2009;29:3991–4001.