Abstract
Fragile X syndrome (FXS) results from the loss of the fragile X mental retardation protein (FMRP), an RNA-binding protein that regulates a variety of cytoplasmic mRNAs. FMRP regulates mRNA translation and may be important in mRNA localization to dendrites. We report a third cytoplasmic regulatory function for FMRP: control of mRNA stability. In mice, we found that FMRP binds, in vivo, the mRNA encoding PSD-95, a key molecule that regulates neuronal synaptic signaling and learning. This interaction occurs through the 3′ untranslated region of the PSD-95 (also known as Dlg4) mRNA, increasing message stability. Moreover, stabilization is further increased by mGluR activation. Although we also found that the PSD-95 mRNA is synaptically localized in vivo, localization occurs independently of FMRP. Through our functional analysis of this FMRP target we provide evidence that dysregulation of mRNA stability may contribute to the cognitive impairments in individuals with FXS.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Bagni, C. & Greenough, W.T. From mRNP trafficking to spine dysmorphogenesis: the roots of fragile X syndrome. Nat. Rev. Neurosci. 6, 376–387 (2005).
Darnell, J.C., Mostovetsky, O. & Darnell, R.B. FMRP RNA targets: identification and validation. Genes Brain Behav. 4, 341–349 (2005).
Zalfa, F., Achsel, T. & Bagni, C. mRNPs, polysomes or granules: FMRP in neuronal protein synthesis. Curr. Opin. Neurobiol. 16, 265–269 (2006).
Darnell, J.C. et al. Fragile X mental retardation protein targets G-quartet mRNAs important for neuronal function. Cell 107, 489–499 (2001).
Schaeffer, C. et al. The fragile X mental retardation protein binds specifically to its mRNA via a purine quartet motif. EMBO J. 20, 4803–4813 (2001).
Ramos, A., Hollingworth, D. & Pastore, A. G-quartet–dependent recognition between the FMRP RGG box and RNA. RNA 9, 1198–1207 (2003).
Chen, L., Yun, S.W., Seto, J., Liu, W. & Toth, M. The fragile X mental retardation protein binds and regulates a novel class of mRNAs containing U-rich target sequences. Neuroscience 120, 1005–1017 (2003).
Zalfa, F. et al. The fragile X syndrome protein FMRP associates with BC1 RNA and regulates the translation of specific mRNAs at synapses. Cell 112, 317–327 (2003).
Gabus, C., Mazroui, R., Tremblay, S., Khandjian, E.W. & Darlix, J.L. The fragile X mental retardation protein has nucleic acid chaperone properties. Nucleic Acids Res. 32, 2129–2137 (2004).
Zalfa, F. et al. Fragile X mental retardation protein (FMRP) binds specifically to the brain cytoplasmic RNAs BC1/BC200 via a novel RNA-binding motif. J. Biol. Chem. 280, 33403–33410 (2005).
Johnson, E.M. et al. Role of Pur alpha in targeting mRNA to sites of translation in hippocampal neuronal dendrites. J. Neurosci. Res. 83, 929–943 (2006).
Jin, P., Alisch, R.S. & Warren, S.T. RNA and microRNAs in fragile X mental retardation. Nat. Cell Biol. 6, 1048–1053 (2004).
Darnell, J.C. et al. Kissing complex RNAs mediate interaction between the fragile X mental retardation protein KH2 domain and brain polyribosomes. Genes Dev. 19, 903–918 (2005).
Kanai, Y., Dohmae, N. & Hirokawa, N. Kinesin transports RNA: isolation and characterization of an RNA-transporting granule. Neuron 43, 513–525 (2004).
Zhong, N., Ju, W., Nelson, D., Dobkin, C. & Brown, W.T. Reduced mRNA for G3BP in fragile X cells: evidence of FMR1 gene regulation. Am. J. Med. Genet. 84, 268–271 (1999).
Brown, V. et al. Microarray identification of FMRP-associated brain mRNAs and altered mRNA translational profiles in fragile X syndrome. Cell 107, 477–487 (2001).
Miyashiro, K.Y. et al. RNA cargoes associating with FMRP reveal deficits in cellular functioning in Fmr1 null mice. Neuron 37, 417–431 (2003).
Huber, K.M., Gallagher, S.M., Warren, S.T. & Bear, M.F. Altered synaptic plasticity in a mouse model of fragile X mental retardation. Proc. Natl. Acad. Sci. USA 99, 7746–7750 (2002).
Tonegawa, S. et al. Hippocampal CA1–region–restricted knockout of NMDAR1 gene disrupts synaptic plasticity, place fields and spatial learning. Cold Spring Harb. Symp. Quant. Biol. 61, 225–238 (1996).
Migaud, M. et al. Enhanced long-term potentiation and impaired learning in mice with mutant postsynaptic density–95 protein. Nature 396, 433–439 (1998).
Fagiolini, M. et al. Separable features of visual cortical plasticity revealed by N-methyl-D-aspartate receptor 2A signaling. Proc. Natl. Acad. Sci. USA 100, 2854–2859 (2003).
Silva, A.J., Paylor, R., Wehner, J.M. & Tonegawa, S. Impaired spatial learning in α-calcium-calmodulin kinase II mutant mice. Science 257, 206–211 (1992).
Husi, H., Ward, M.A., Choudhary, J.S., Blackstock, W.P. & Grant, S.G. Proteomic analysis of NMDA receptor–adhesion protein signaling complexes. Nat. Neurosci. 3, 661–669 (2000).
Sheng, M. & Kim, M.J. Postsynaptic signaling and plasticity mechanisms. Science 298, 776–780 (2002).
Cuthbert, P.C. et al. SAP102/dlgh3 couples the NMDA receptor to specific plasticity pathways and learning strategies. J. Neurosci. 27, 2673–2682 (2007).
Tarpey, P. et al. Mutations in the DLG3 gene cause nonsyndromic X-linked mental retardation. Am. J. Hum. Genet. 75, 318–324 (2004).
Vickers, C.A. et al. Neurone specific regulation of dendritic spines in vivo by post synaptic density–95 protein (PSD-95). Brain Res (2006).
Reiss, A.L., Lee, J. & Freund, L. Neuroanatomy of fragile X syndrome: the temporal lobe. Neurology 44, 1317–1324 (1994).
Todd, P.K., Mack, K.J. & Malter, J.S. The fragile X mental retardation protein is required for type-I metabotropic glutamate receptor–dependent translation of PSD-95. Proc. Natl. Acad. Sci. USA 100, 14374–14378 (2003).
Zhang, Y.Q. et al. Drosophila fragile X–related gene regulates the MAP1B homolog Futsch to control synaptic structure and function. Cell 107, 591–603 (2001).
Lu, R. et al. The fragile X protein controls microtubule-associated protein 1B translation and microtubule stability in brain neuron development. Proc. Natl. Acad. Sci. USA 101, 15201–15206 (2004).
Niranjanakumari, S., Lasda, E., Brazas, R. & Garcia-Blanco, M.A. Reversible crosslinking combined with immunoprecipitation to study RNA-protein interactions in vivo. Methods 26, 182–190 (2002).
Bence, M., Arbuckle, M.I., Dickson, K.S. & Grant, S.G. Analyses of murine postsynaptic density–95 identify novel isoforms and potential translational control elements. Brain Res. Mol. Brain Res. 133, 143–152 (2005).
Adinolfi, S. et al. Dissecting FMR1, the protein responsible for fragile X syndrome, in its structural and functional domains. RNA 5, 1248–1258 (1999).
Williamson, J.R., Raghuraman, M.K. & Cech, T.R. Monovalent cation–induced structure of telomeric DNA: the G-quartet model. Cell 59, 871–880 (1989).
Klann, E. & Dever, T.E. Biochemical mechanisms for translational regulation in synaptic plasticity. Nat. Rev. Neurosci. 5, 931–942 (2004).
Pillai, R.S., Bhattacharyya, S.N. & Filipowicz, W. Repression of protein synthesis by miRNAs: how many mechanisms? Trends Cell Biol. 17, 118–126 (2007).
Steward, O. & Schuman, E.M. Compartmentalized synthesis and degradation of proteins in neurons. Neuron 40, 347–359 (2003).
Lein, E.S. et al. Genome-wide atlas of gene expression in the adult mouse brain. Nature 445, 168–176 (2007).
Loesch, D.Z., Huggins, R.M. & Hagerman, R.J. Phenotypic variation and FMRP levels in fragile X. Ment. Retard. Dev. Disabil. Res. Rev. 10, 31–41 (2004).
Wang, H. et al. Developmentally-programmed FMRP expression in oligodendrocytes: a potential role of FMRP in regulating translation in oligodendroglia progenitors. Hum. Mol. Genet. 13, 79–89 (2003).
Weiler, I.J. et al. Fragile X mental retardation protein is translated near synapses in response to neurotransmitter activation. Proc. Natl. Acad. Sci. USA 94, 5395–5400 (1997).
Antar, L.N., Afroz, R., Dictenberg, J.B., Carroll, R.C. & Bassell, G.J. Metabotropic glutamate receptor activation regulates fragile X mental retardation protein and FMR1 mRNA localization differentially in dendrites and at synapses. J. Neurosci. 24, 2648–2655 (2004).
Barreau, C., Paillard, L. & Osborne, H.B. AU-rich elements and associated factors: are there unifying principles? Nucleic Acids Res. 33, 7138–7150 (2005).
Smith, C.L. et al. GAP-43 mRNA in growth cones is associated with HuD and ribosomes. J. Neurobiol. 61, 222–235 (2004).
Singh, R. & Valcarcel, J. Building specificity with nonspecific RNA-binding proteins. Nat. Struct. Mol. Biol. 12, 645–653 (2005).
Ule, J. & Darnell, R.B. RNA binding proteins and the regulation of neuronal synaptic plasticity. Curr. Opin. Neurobiol. 16, 102–110 (2006).
Plath, N. et al. Arc/Arg3.1 is essential for the consolidation of synaptic plasticity and memories. Neuron 52, 437–444 (2006).
Gylys, K.H. et al. Synaptic changes in Alzheimer's disease: increased amyloid-beta and gliosis in surviving terminals is accompanied by decreased PSD-95 fluorescence. Am. J. Pathol. 165, 1809–1817 (2004).
Toro, C. & Deakin, J.F. NMDA receptor subunit NRI and postsynaptic protein PSD-95 in hippocampus and orbitofrontal cortex in schizophrenia and mood disorder. Schizophr. Res. 80, 323–330 (2005).
Acknowledgements
We thank B.A. Oostra for the FMR1 knockout mice, N.K. Gray and T. Achsel for their critical evaluation of the manuscript, and O. Steward for precious suggestions and reagents. We thank M.A. Kiebler for advice on the neuronal transfection protocol. This research was funded by a European Molecular Biology Organization short-term fellowship, a Royal Society of Edinburgh Scottish Executive Enterprise and Lifelong Learning Department fellowship and a Biotechnology and Biological Sciences Research Council grant (C19143) to K.S.D., by Telethon (GGP05269), Ministero della Salute, Ministero della Università (FIRB) to C.B. and by Wellcome Trust grant number 056523 and the Wellcome Trust Genes to Cognition Programe to S.G.N.G. F.Z. was supported by the Associazione Italiana Sindrome dell'X Fragile.
Author information
Authors and Affiliations
Contributions
F.Z. contributed to the conclusions drawn in Figures 1,4,6,7 and 8. B.E. contributed to the conclusions drawn from Figures 5,6,7,8. K.S.D. provided intellectual input, contributed to the conclusions drawn from Figures 1,2,3 and 8, contributed a portion of the funding and contributed to the writing of this manuscript. V.M. contributed to the conclusions drawn from Figures 5,6,7. S.D.R. contributed to the conclusions drawn from Figures 2,3 and 7. A.D.P. contributed to the conclusions drawn from Figures 1 and 7. E.T. and P.C. contributed to the conclusions drawn from Figures 7 and 8. G.N. contributed with intellectual inputs. S.G.N.G. contributed some initial funding for this work and intellectual inputs. C.B. provided intellectual input, funding, coordination of the project and contributed to the writing of this manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Fig. 1
G-quartet consensus and similarities to PSD-95 mRNA. (PDF 710 kb)
Supplementary Fig. 2
PSD-95 mRNA localizes in synaptoneurosomes from total brain. (PDF 964 kb)
Supplementary Fig. 3
PSD-95 mRNA is specifically detected by in situ hybridization. (PDF 1507 kb)
Supplementary Fig. 4
PSD-95 mRNA is dendritcally localized in vivo. (PDF 752 kb)
Supplementary Fig. 5
FMRP regulates the stability of PSD-95 mRNA in hippocampal cells. (PDF 319 kb)
Supplementary Fig. 6
FMRP doesn't regulate the stability of PSD-95 mRNA in cortical cells. (PDF 275 kb)
Supplementary Fig. 7
Viability of WT and FMR1 KO hippocampal neurons during actinomycin D treatments. (PDF 731 kb)
Supplementary Fig. 8
HuD and HuR protein levels in mouse hippocampus, cerebellum and cortex. (PDF 421 kb)
Supplementary Table 1
Stability assay of other FMRP target mRNAs. (PDF 65 kb)
Rights and permissions
About this article
Cite this article
Zalfa, F., Eleuteri, B., Dickson, K. et al. A new function for the fragile X mental retardation protein in regulation of PSD-95 mRNA stability. Nat Neurosci 10, 578–587 (2007). https://doi.org/10.1038/nn1893
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nn1893
This article is cited by
-
FMRP ligand circZNF609 destabilizes RAC1 mRNA to reduce metastasis in acral melanoma and cutaneous melanoma
Journal of Experimental & Clinical Cancer Research (2022)
-
NMD abnormalities during brain development in the Fmr1-knockout mouse model of fragile X syndrome
Genome Biology (2021)
-
FMRP regulates STAT3 mRNA localization to cellular protrusions and local translation to promote hepatocellular carcinoma metastasis
Communications Biology (2021)
-
Fragile X mental retardation protein in intrahepatic cholangiocarcinoma: regulating the cancer cell behavior plasticity at the leading edge
Oncogene (2021)
-
The molecular biology of FMRP: new insights into fragile X syndrome
Nature Reviews Neuroscience (2021)