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Dynamic O-GlcNAc modification regulates CREB-mediated gene expression and memory formation

Nature Chemical Biology volume 8, pages 253–261 (2012)Cite this article

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Abstract

The transcription factor cyclic AMP–response element binding protein (CREB) is a key regulator of many neuronal processes, including brain development, circadian rhythm and long-term memory. Studies of CREB have focused on its phosphorylation, although the diversity of CREB functions in the brain suggests additional forms of regulation. Here we expand on a chemoenzymatic strategy for quantifying glycosylation stoichiometries to characterize the functional roles of CREB glycosylation in neurons. We show that CREB is dynamically modified with an O-linked β-N-acetyl-D-glucosamine sugar in response to neuronal activity and that glycosylation represses CREB-dependent transcription by impairing its association with CREB-regulated transcription coactivator (CRTC; also known as transducer of regulated CREB activity). Blocking glycosylation of CREB alters cellular function and behavioral plasticity, enhancing both axonal and dendritic growth and long-term memory consolidation. Our findings demonstrate a new role for O-glycosylation in memory formation and provide a mechanistic understanding of how glycosylation contributes to critical neuronal functions. Moreover, we identify a previously unknown mechanism for the regulation of activity-dependent gene expression, neural development and memory.

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Figure 1: CREB is O-GlcNAc glycosylated at Ser40 in response to neuronal activity.
Figure 2: Neuronal activity induces CREB glycosylation preferentially on the Ser133-phosphorylated subpopulation.
Figure 3: Glycosylation at Ser40 represses CREB activity via a CRTC-dependent mechanism.
Figure 4: CREB glycosylation at Ser40 represses dendritic and axonal growth.
Figure 5: CREB glycosylation at Ser40 modulates long-term conditioned fear memory.

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Acknowledgements

We thank M. Antoniou (King's College London School of Medicine) for the pA2UCOE-EGFP construct, M. Greenberg (Harvard University) for the pLEMPRA-GOI and pLLX-shRNA constructs, G. Hart (The John Hopkins University School of Medicine) for the OGT-specific antibody, S. Josselyn (University of Toronto) for the p1005-CREB construct, R. Lansford (California Institute of Technology) for the pLenti PGK-H2B-mCherry construct, R. Malenka (Stanford University) and X. Yu (Shanghai Institutes for Biological Sciences) for the pcDNA3-Dkk-1-Flag and Ncad(intra) constructs, P. Qasba (US National Cancer Institute) for the Y289L GalT construct and L. Wells (University of Georgia) for the pDEST-HA-OGA construct. We thank S.-H. Yu (California Institute of Technology) for synthesizing the UDP-ketogalactose substrate and D. Anderson (California Institute of Technology) for providing the fear conditioning apparatus. We thank A. Silva for a critical reading of the manuscript. This work was supported by grants from the US National Institutes of Health (R01 GM084724 to L.C.H.-W., F31 NS056525 to J.E.R. and National Research Service Award Training Grant 5T32 GM07737 to P.M.C.).

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  1. Jessica E Rexach and Peter M Clark: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, USA

    Jessica E Rexach, Peter M Clark & Linda C Hsieh-Wilson

  2. Howard Hughes Medical Institute, California Institute of Technology, Pasadena, California, USA

    Jessica E Rexach, Peter M Clark & Linda C Hsieh-Wilson

  3. Genomics Institute of the Novartis Research Foundation, San Diego, California, USA

    Daniel E Mason & Eric C Peters

  4. Picower Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Rachael L Neve

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Contributions

L.C.H.-W. designed, directed and coordinated the project. P.M.C. and J.E.R. designed and performed the experiments except where otherwise noted. D.E.M. and E.C.P. performed the MS analyses; R.L.N. prepared the HSV. P.M.C., J.E.R. and L.C.H.-W. wrote the manuscript, and all authors participated in editing it.

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Correspondence to Linda C Hsieh-Wilson.

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Rexach, J., Clark, P., Mason, D. et al. Dynamic O-GlcNAc modification regulates CREB-mediated gene expression and memory formation. Nat Chem Biol 8, 253–261 (2012). https://doi.org/10.1038/nchembio.770

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