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A new function for the fragile X mental retardation protein in regulation of PSD-95 mRNA stability

Nature Neuroscience volume 10, pages 578–587 (2007)Cite this article

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.

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Figure 1: FMRP interacts directly with the 3′ UTR of PSD-95 mRNA.
Figure 2: The C-terminal domain of FMRP is able to specifically interact with the PSD-95 mRNA 3′ UTR.
Figure 3: A G-rich region in the PSD-95 3′ UTR is responsible for FMRP C-terminus binding.
Figure 4: The PSD-95 mRNA polysomal profile is similar in wild-type and FMR1 knockout mice.
Figure 5: PSD-95 mRNA is dendritically localized in neuronal cell cultures.
Figure 6: PSD-95 mRNA is dendritically localized in vivo.
Figure 7: PSD-95 mRNA and protein levels are altered in the FMR1 knockout mice.
Figure 8: FMRP regulates the stability of PSD-95 mRNA in hippocampal cells through an activity-dependent mechanism.

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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

Author notes

  1. Francesca Zalfa, Boris Eleuteri and Kirsten S Dickson: These authors contributed equally to this work.

Authors and Affiliations

  1. Dipartimento di Biologia, Università “Tor Vergata”, Via della Ricerca Scientifica 1, Rome, 00133, Italy

    Francesca Zalfa, Boris Eleuteri, Valentina Mercaldo, Silvia De Rubeis & Claudia Bagni

  2. Istituto di Neuroscienze Sperimentali, Fondazione Santa Lucia, Via del Fosso di Fiorano 63, Rome, 00143, Italy

    Francesca Zalfa, Boris Eleuteri, Valentina Mercaldo, Silvia De Rubeis, Alessandra di Penta & Claudia Bagni

  3. Division of Neuroscience, University of Edinburgh, George Sq., Edinburgh, EH8 9JZ, UK

    Kirsten S Dickson & Seth G N Grant

  4. Istituto di Genetica Medica, Università Cattolica, Largo F. Vito 1, Rome, 00168, Italy

    Elisabetta Tabolacci, Pietro Chiurazzi & Giovanni Neri

  5. Wellcome Trust Sanger Institute, Hinxton, Cambridgeshire, CB10 1SA, UK

    Seth G N Grant

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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

Correspondence to Kirsten S Dickson or Claudia Bagni.

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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)

Supplementary Methods (PDF 145 kb)

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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

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