C-terminal interactors of the AMPA receptor auxiliary subunit Shisa9

Anna R Karataeva; Remco V Klaassen; Jasper Ströder; Marta Ruiperez-Alonso; Johannes J J Hjorth; Pim van Nierop; Sabine Spijker; Huibert D Mansvelder; August B Smit

doi:10.1371/journal.pone.0087360

C-terminal interactors of the AMPA receptor auxiliary subunit Shisa9

PLoS One. 2014 Feb 3;9(2):e87360. doi: 10.1371/journal.pone.0087360. eCollection 2014.

Authors

Anna R Karataeva¹, Remco V Klaassen¹, Jasper Ströder², Marta Ruiperez-Alonso³, Johannes J J Hjorth³, Pim van Nierop¹, Sabine Spijker¹, Huibert D Mansvelder³, August B Smit¹

Affiliations

¹ Department of Molecular and Cellular Neurobiology, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, Vrije Universiteit University, Amsterdam, The Netherlands.
² Department of Molecular and Cellular Neurobiology, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, Vrije Universiteit University, Amsterdam, The Netherlands ; Department of Integrative Neurophysiology, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, Vrije Universiteit University, Amsterdam, The Netherlands.
³ Department of Integrative Neurophysiology, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, Vrije Universiteit University, Amsterdam, The Netherlands.

Abstract

Shisa9 (initially named CKAMP44) has been identified as auxiliary subunit of the AMPA-type glutamate receptors and was shown to modulate its physiological properties. Shisa9 is a type-I transmembrane protein and contains a C-terminal PDZ domain that potentially interacts with cytosolic proteins. In this study, we performed a yeast two-hybrid screening that yielded eight PDZ domain-containing interactors of Shisa9, which were independently validated. The identified interactors are known scaffolding proteins residing in the neuronal postsynaptic density. To test whether C-terminal scaffolding interactions of Shisa9 affect synaptic AMPA receptor function in the hippocampus, we disrupted these interactions using a Shisa9 C-terminal mimetic peptide. In the absence of scaffolding interactions of Shisa9, glutamatergic AMPA receptor-mediated synaptic currents in the lateral perforant path of the mouse hippocampus had a faster decay time, and paired-pulse facilitation was reduced. Furthermore, disruption of the PDZ interactions between Shisa9 and its binding partners affected hippocampal network activity. Taken together, our data identifies novel interaction partners of Shisa9, and shows that the C-terminal interactions of Shisa9 through its PDZ domain interaction motif are important for AMPA receptor synaptic and network functions.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs*
Amino Acid Sequence
Animals
Binding Sites / genetics
Blotting, Western
HEK293 Cells
Hippocampus / metabolism
Hippocampus / physiology
Humans
Immunoprecipitation
Mice
Mice, Inbred C57BL
Molecular Sequence Data
Nerve Net / physiology
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism*
Neuronal Plasticity / physiology
Peptides / genetics
Peptides / metabolism*
Protein Binding
Protein Subunits / genetics
Protein Subunits / metabolism
Receptors, AMPA / genetics
Receptors, AMPA / metabolism*
Synapses / metabolism
Synapses / physiology
Synaptic Transmission / physiology
Two-Hybrid System Techniques

Substances

CKAMP44 protein, mouse
Nerve Tissue Proteins
Peptides
Protein Subunits
Receptors, AMPA

Grants and funding

ABS, HDM and AK received funding from the EU 7th Framework Programme (HEALTH-F2-2009-242167 ‘SynSys’). ABS, SS and HDM received funding from the Dutch Fund for Economic Structure Reinforcement (FES, 0908 “NeuroBasic PharmaPhenomics project”) and HDM received funding from the ERC StG ŒBrainSignals). RVK received funding from NWO-ALW (ALW2PJ/12048). JS received funding from the EU 7th Framework ITN 238686 ‘CEREBNET’. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.