Analysis of SUMO1-conjugation at Synapses

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Abstract

SUMO1-conjugation of proteins at neuronal synapses is considered to be a major post-translational regulatory process in nerve cell and synapse function, but the published evidence for SUMO1-conjugation at synapses is contradictory. We employed multiple genetic mouse models for stringently controlled biochemical and immunostaining analyses of synaptic SUMO1-conjugation. By using a knock-in reporter mouse line expressing tagged SUMO1, we could not detect SUMO1-conjugation of seven previously proposed synaptic SUMO1-targets in the brain. Further, immunostaining of cultured neurons from wild-type and SUMO1 knock-out mice showed that anti-SUMO1 immunolabelling at synapses is non-specific. Our findings indicate that SUMO1-conjugation of synaptic proteins does not occur or is extremely rare and hence not detectable using current methodology. Based on our data, we discuss a set of experimental strategies and minimal consensus criteria for the validation of SUMOylation that can be applied to any SUMOylation substrate and SUMO isoform.

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James A Daniel

Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, Germany

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-2781-4544
Benjamin H Cooper

Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, Germany

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The authors declare that no competing interests exist.
Jorma J Palvimo

Institute of Biomedicine, University of Eastern Finland, Kuopio, Finland

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The authors declare that no competing interests exist.
Fu-ping Zhang

Institute of Biomedicine, University of Turku, Turku, Finland

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The authors declare that no competing interests exist.
Nils Brose

Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, Germany

Competing interests
The authors declare that no competing interests exist.
Marilyn Tirard

Department of Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, Germany

For correspondence
tirard@em.mpg.de

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-5669-9610

Funding

Max-Planck-Gesellschaft (Open-access funding)

Marilyn Tirard

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments were performed in accordance with the guidelines for the welfare of experimental animals issued by the State Government of Lower Saxony, Germany, in compliance with European and NIH guidelines (33.9-42502-04-13/1359).

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This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.