Synaptic Strength Regulated by Palmitate Cycling on PSD-95

El‐Husseini, Alaa; Schnell, Eric; Dakoji, Srikanth; Sweeney, Neal; Zhou, Qiang; Prange, Oliver; Gauthier-Campbell, Catherine; Aguilera-Moreno, Andrea; Nicoll, Roger A.; Bredt, David S.

doi:10.1016/s0092-8674(02)00683-9

Cited by 517 publications

(461 citation statements)

References 57 publications

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“…Next, we fused the prenylated and dual palmitoylated motif of paralemmin to the COOH-terminus of JNK3 CS to mimic a constant pseudo-palmitoylation on JNK3 (JNK3 Parlm). [34][35][36] The effects of Wnt7a on axonal branching were completely blocked in neurons overexpressing JNK3 Parlm (secondary, 2.8±0.2, Po0.01; tertiary, 0.5±0.1, Po0.01; higher, 0.0 ± 0.0, Po0.01; total, 4.3 ± 0.3, Po0.01) compared with WT treated with Wnt7a (Figures 4c and d), suggesting that it is in fact JNK3 palmitoylation that modulates Wnt7a-induced axonal branching. Furthermore, overexpression of JNK3 Parlm was found to reduce axonal branching and total axon length in untreated neurons, compared with controls ( Figure 4e).…”

Section: Resultsmentioning

confidence: 99%

Isoform-specific palmitoylation of JNK regulates axonal development

et al. 2011

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The c-jun N-terminal kinase (JNK) proteins are encoded by three genes (Jnk1-3), giving rise to 10 isoforms in the mammalian brain. The differential roles of JNK isoforms in neuronal cell death and development have been noticed in several pathological and physiological contexts. However, the mechanisms underlying the regulation of different JNK isoforms to fulfill their specific roles are poorly understood. Here, we report an isoform-specific regulation of JNK3 by palmitoylation, a posttranslational modification, and the involvement of JNK3 palmitoylation in axonal development and morphogenesis. Two cysteine residues at the COOH-terminus of JNK3 are required for dynamic palmitoylation, which regulates JNK3's distribution on the actin cytoskeleton. Expression of palmitoylation-deficient JNK3 increases axonal branching and the motility of axonal filopodia in cultured hippocampal neurons. The Wnt family member Wnt7a, a known modulator of axonal branching and remodelling, regulates the palmitoylation and distribution of JNK3. Palmitoylation-deficient JNK3 mimics the effect of Wnt7a application on axonal branching, whereas constitutively palmitoylated JNK3 results in reduced axonal branches and blocked Wnt7a induction. Our results demonstrate that protein palmitoylation is a novel mechanism for isoform-specific regulation of JNK3 and suggests a potential role of JNK3 palmitoylation in modulating axonal branching. The c-jun N-terminal kinase (JNK) family consists of JNK1, JNK2 and JNK3 subgroups, which participate in diverse biological and pathological processes. 1,2 At least 10 JNK isoforms of varied sizes, with most between 46 and 54 kDa, are produced from the Jnk1-3 genes, giving rise to JNK isoforms. 1 In the nervous system, JNKs (particularly isoform JNK3) have been extensively studied as the key players in apoptosis and neurodegeneration. [3][4][5][6] However, accumulating evidence supports a physiological role of JNK in regulating neurite formation and morphogenesis. 7-12 Pharmacological inhibition of JNK activity blocks axogenesis in hippocampal neurons, arguing for an essential role of JNK in neurite development. 13 Growth factors and signalling molecules, including secreted proteins of the Wnt family, have also been found to activate JNK for remodelling dendrites and axons, a process that relies on cytoskeletal rearrangement. 11,[14][15][16] This has led to the identification of several actin-or microtubule-associated proteins as JNK substrates that modulate different aspects of cytoskeletal activity. 17 However, all the 10 JNK isoforms share the same kinase domain and activation mechanism. 17 The differential roles of JNK isoforms in neurite development and the mechanisms underlying isoform-specific regulation are poorly understood.Analysis of animals null for particular JNK isoforms provides the first evidence to support isoform-specific roles of JNKs in the brain. Mice with Jnk1 À/À show abnormalities in neurite development, 7 whereas mice null for Jnk1 and Jnk2 show embryonic lethality due to severe neuro...

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Section: Resultsmentioning

confidence: 99%

Isoform-specific palmitoylation of JNK regulates axonal development

et al. 2011

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“…First, two-dimensional gel experiments reveal that many distinct proteins are detected as multiple protein spots, suggesting that they exist as isoforms or are post-translationally modified. Phosphorylation and to a lesser extent palmitoylation and nitrosylation of a number of PSD proteins have been implicated in the regulation of the proteins' intrinsic properties and/or their trafficking and localization, and play important roles in the induction and expression of synaptic plasticity (31)(32)(33). Second, the PSD contains distinct functional groups of proteins that might be involved in diverse physiological activities of the PSD.…”

Section: Discussionmentioning

confidence: 99%

Proteomics Analysis of Rat Brain Postsynaptic Density

Hornshaw

Schors

et al. 2004

Journal of Biological Chemistry

235

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The postsynaptic density contains multiple protein complexes that together relay the presynaptic neurotransmitter input to the activation of the postsynaptic neuron. In the present study we took two independent proteome approaches for the characterization of the protein complement of the postsynaptic density, namely 1) two-dimensional gel electrophoresis separation of proteins in conjunction with mass spectrometry to identify the tryptic peptides of the protein spots and 2) isolation of the trypsin-digested sample that was labeled with isotope-coded affinity tag, followed by liquid chromatography-tandem mass spectrometry for the partial separation and identification of the peptides, respectively. Functional grouping of the identified proteins indicates that the postsynaptic density is a structurally and functionally complex organelle that may be involved in a broad range of synaptic activities. These proteins include the receptors and ion channels for glutamate neurotransmission, proteins for maintenance and modulation of synaptic architecture, sorting and trafficking of membrane proteins, generation of anaerobic energy, scaffolding and signaling, local protein synthesis, and correct protein folding and breakdown of synaptic proteins. Together, these results imply that the postsynaptic density may have the ability to function (semi-) autonomously and may direct various cellular functions in order to integrate synaptic physiology.The majority of excitatory neurotransmission in the brain occurs via glutamatergic synapses. In the presynaptic element of the synapse, specialized secretion machinery determines the activity-dependent membrane fusion of glutamate-containing vesicles and the release of transmitter into the synaptic cleft. In the postsynaptic element, glutamate receptors and downstream signal transduction are organized by the protein assembly of the postsynaptic density (PSD). 1 Both the presynaptic release machinery and the PSD are electron-dense assemblies (1, 2), in which proteins are thought to be organized into distinct functional complexes (3-5) that may be dynamically regulated by neuronal activity (6 -8). The modulation of this molecular architecture of the synapse is at the basis of synaptic plasticity (6 -8), and aberrations thereof may underlie neuronal disorders.In view of the importance of the PSD in glutamatergic neurotransmission and its involvement in neuroplasticity, considerable efforts have been made to identify its protein constituents as a prelude to understand the molecular basis of PSD functioning. In the past several years, yeast two-hybrid technology has been extensively used to characterize proteins that interact with the glutamate receptors and may constitute core elements of the PSD involved in the regulation of receptor trafficking and signaling (reviewed in Refs. 9 -11). Based largely on these studies a protein-protein interaction map of the PSD has emerged. In brief, the model posits that the postsynaptic receptor complexes are localized by scaffolding proteins such as the s...

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“…The validity of this compound as an inhibitor of protein palmitoylation has been verified for at least 2 dozen palmitoylated proteins, including Src family kinases, Rho family proteins, H-Ras, PSD95 and transmembrane receptors such as the Nicotinic α7 Receptor and CCR5 [19][20][21][22][23][24]. The advantages of this approach are threefold: 1) the reagent is simple and inexpensive to use; 2) the function of endogenous YFPP can be studied without the need to generate and overexpress mutants; 3) the modified cysteine residue is left intact.…”

Section: Inhibition Of Protein Palmitoylation With 2-bromopalmitate (mentioning

confidence: 99%

Use of analogs and inhibitors to study the functional significance of protein palmitoylation

Resh

2006

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Covalent attachment of palmitate to proteins is a post-translational modification that exerts diverse effects on protein localization and function. The three key technical approaches required for an investigator to determine the role of palmitoylation of Your Favorite Palmitoylated Protein (YFPP) are methods to: 1) detect YFPP palmitoylation; 2) alter or inhibit palmitoylation of YFPP; 3) determine the functional significance of altered YFPP palmitoylation. Here, I describe experimental methods to address these three issues. Both radioactive (radiolabeling with 3 H palmitate or 125 I-IC16 palmitate) and non-radioactive (chemical labeling and mass spectrometry) methods to detect palmitoylated proteins are presented. Next, techniques to inhibit protein palmitoylation are described. These include site specific mutagenesis, and treatment of cells with inhibitors of protein palmitoylation, including 2-bromopalmitate, cerulenin, and tunicamycin. Alternative methods to replace palmitate with other fatty acids are also presented. Finally, general approaches to determining the effect of altered palmitoylation status on YFPP association with membranes and lipid rafts, as well as signal transduction, are described.

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Synaptic Strength Regulated by Palmitate Cycling on PSD-95

Cited by 517 publications

References 57 publications

Isoform-specific palmitoylation of JNK regulates axonal development

Isoform-specific palmitoylation of JNK regulates axonal development

Proteomics Analysis of Rat Brain Postsynaptic Density

Use of analogs and inhibitors to study the functional significance of protein palmitoylation

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