The goals of the proposed investigation are to study the regulation of the tyrosine phosphatase STEP. Previous studies established that STEP inactivates ERK1/2, p38 and the tyrosine kinase Fyn. Over the last cycle, we have discovered that STEP also induces NR1/NR2B and GluR1/GluR2 internalization. STEP has been implicated in the pathophysiology of Fragile X syndrome (FXS), Alzheimer's disease (AD), and schizophrenia (SZ). In these disorders, there is an increase in STEP levels. The current model suggests that the increase in STEP leads to the inappropriate internalization of NR1/NR2B and GluR1/GluR2 receptors. However, the mechanisms by which STEP levels are increased differ in these disorders. In AD and SZ, there is a disruption in the ubiquitination and degradation STEP. In FXS, there is an increase in the translation of STEP mRNA due to the absence of FMRP. STEP is rapidly degraded after specific forms of synaptic activity. Synaptic stimulation results in the ubiquitination of STEP, which is then degraded through the proteasome system. We hypothesize that STEP is degraded at synaptic sites to promote synaptic strengthening. Extrasynaptic NMDAR stimulation results in a calpain-mediated proteolysis STEP. We propose that this mechanism prolongs p38 signaling and initiates cell death pathways. The molecular mechanisms that regulate these processes are unknown.
Aim 1 focuses on STEP ubiquitination. We include functional experiments that mutate STEP so that it can no longer be ubiquitinated and degraded. We predict that this construct will disrupt both LTP, as well as the consolidation of fear conditioning after infusion into the amygdala. We will determine the kinases that promote or prevent STEP ubiquitination.
Aim 2 focuses on STEP proteolysis. We include functional studies that disrupt proteolysis of STEP, and will determine the effects this has on cell death using in vivo excitotoxic models. We predict that blocking STEP proteolysis will significantly decrease cell death.
Aim 3 focuses on STEP translation. Two cytoplasmic polyadenylation elements (CPEs) are present in STEP mRNA and we will determine whether these elements regulate the translation of STEP mRNA. We include functional experiments that will mutate these CPEs. We predict this will disrupt the normal function of STEP in the regulation of its substrates, including glutamate receptor trafficking. These studies will advance our understanding of how STEP is involved in normal cognitive processes and possible mechanisms by which it is disrupted in several diseases in which there are significant cognitive deficits.

Public Health Relevance

A major discovery over the last cycle is that STriatal-Enriched tyrosine Phosphatase (STEP) regulates glutamate receptor trafficking, and removes both NR1/NR2B and GluR1/GluR2 receptors from synaptic membranes after specific forms of synaptic stimulation. These latter findings have implicated STEP in the pathophysiology of fragile X syndrome, schizophrenia, and Alzheimer's disease. The proposed studies examine the molecular mechanisms that regulate STEP activity and will clarify how this important family of regulatory proteins are involved in synaptic plasticity, cell death, and human disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH052711-20
Application #
8644881
Study Section
Molecular Neuropharmacology and Signaling Study Section (MNPS)
Program Officer
Asanuma, Chiiko
Project Start
1999-05-01
Project End
2015-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
20
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Yale University
Department
Psychiatry
Type
Schools of Medicine
DUNS #
City
New Haven
State
CT
Country
United States
Zip Code
06510
Xu, J; Hartley, B J; Kurup, P et al. (2016) Inhibition of STEP61 ameliorates deficits in mouse and hiPSC-based schizophrenia models. Mol Psychiatry :
Saavedra, Ana; Puigdellívol, Mar; Tyebji, Shiraz et al. (2016) BDNF Induces Striatal-Enriched Protein Tyrosine Phosphatase 61 Degradation Through the Proteasome. Mol Neurobiol 53:4261-73
Xu, Jian; Kurup, Pradeep; Azkona, Garikoitz et al. (2016) Down-regulation of BDNF in cell and animal models increases striatal-enriched protein tyrosine phosphatase 61 (STEP61 ) levels. J Neurochem 136:285-94
Azkona, Garikoitz; Saavedra, Ana; Aira, Zigor et al. (2016) Striatal-enriched protein tyrosine phosphatase modulates nociception: evidence from genetic deletion and pharmacological inhibition. Pain 157:377-86
Jang, Sung-Soo; Royston, Sara E; Xu, Jian et al. (2015) Regulation of STEP61 and tyrosine-phosphorylation of NMDA and AMPA receptors during homeostatic synaptic plasticity. Mol Brain 8:55
Kurup, Pradeep K; Xu, Jian; Videira, Rita Alexandra et al. (2015) STEP61 is a substrate of the E3 ligase parkin and is upregulated in Parkinson's disease. Proc Natl Acad Sci U S A 112:1202-7
Legastelois, Rémi; Darcq, Emmanuel; Wegner, Scott A et al. (2015) Striatal-enriched protein tyrosine phosphatase controls responses to aversive stimuli: implication for ethanol drinking. PLoS One 10:e0127408
Xu, Jian; Kurup, Pradeep; Foscue, Ethan et al. (2015) Striatal-enriched protein tyrosine phosphatase regulates the PTPα/Fyn signaling pathway. J Neurochem 134:629-41
Chiodi, Valentina; Mallozzi, Cinzia; Ferrante, Antonella et al. (2014) Cocaine-induced changes of synaptic transmission in the striatum are modulated by adenosine A2A receptors and involve the tyrosine phosphatase STEP. Neuropsychopharmacology 39:569-78
Darcq, Emmanuel; Hamida, Sami Ben; Wu, Su et al. (2014) Inhibition of striatal-enriched tyrosine phosphatase 61 in the dorsomedial striatum is sufficient to increased ethanol consumption. J Neurochem 129:1024-34

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