University of Oulu

Eriksen, M.S., Nikolaienko, O., Hallin, E.I., Grødem, S., Bustad, H.J., Flydal, M.I., Merski, I., Hosokawa, T., Lascu, D., Akerkar, S., Cuéllar, J., Chambers, J.J., O’Connell, R., Muruganandam, G., Loris, R., Touma, C., Kanhema, T., Hayashi, Y., Stratton, M.M., Valpuesta, J.M., Kursula, P., Martinez, A. and Bramham, C.R. (2021), Arc self-association and formation of virus-like capsids are mediated by an N-terminal helical coil motif. FEBS J, 288: 2930-2955. https://doi.org/10.1111/febs.15618

Arc self-association and formation of virus-like capsids are mediated by an N-terminal helical coil motif

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Author: Eriksen, Maria S.1,2; Nikolaienko, Oleksii1,2; Hallin, Erik I.1;
Organizations: 1Department of Biomedicine, University of Bergen, Norway
2KG Jebsen Centre for Neuropsychiatric Disorders, University of Bergen, Norway
3Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, MA, USA
4Department of Pharmacology, Kyoto University Graduate School of Medicine, Kyoto, Japan
5Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain
6Institute for Applied Life Sciences, University of Massachusetts Amherst, MA, USA
7VIB-VUB Center for Structural Biology, Vlaams Instituut voor Biotechnologie, Brussels, Belgium
8Structural Biology Brussels, Department of Bioengineering Sciences, Vrije Universiteit Brussel, Belgium
9Faculty of Biochemistry and Molecular Biology & Biocenter Oulu, University of Oulu, Finland
Format: article
Version: accepted version
Access: embargoed
Persistent link: http://urn.fi/urn:nbn:fi-fe2021070140815
Language: English
Published: John Wiley & Sons, 2021
Publish Date: 2021-11-11
Description:

Abstract

Activity-regulated cytoskeleton-associated protein (Arc) is a protein interaction hub with diverse roles in intracellular neuronal signaling, and important functions in neuronal synaptic plasticity, memory, and postnatal cortical development. Arc has homology to retroviral Gag protein and is capable of self-assembly into virus-like capsids implicated in the intercellular transfer of RNA. However, the molecular basis of Arc self-association and capsid formation is largely unknown. Here, we identified a 28-amino-acid stretch in the mammalian Arc N-terminal (NT) domain that is necessary and sufficient for self-association. Within this region, we identified a 7-residue oligomerization motif, critical for the formation of virus-like capsids. Purified wild-type Arc formed capsids as shown by transmission and cryo-electron microscopy, whereas mutant Arc with disruption of the oligomerization motif formed homogenous dimers. An atomic-resolution crystal structure of the oligomerization region peptide demonstrated an antiparallel coiled-coil interface, strongly supporting NT-NT domain interactions in Arc oligomerization. The NT coil–coil interaction was also validated in live neurons using fluorescence lifetime FRET imaging, and mutation of the oligomerization motif disrupted Arc-facilitated endocytosis. Furthermore, using single-molecule photobleaching, we show that Arc mRNA greatly enhances higher-order oligomerization in a manner dependent on the oligomerization motif. In conclusion, a helical coil in the Arc NT domain supports self-association above the dimer stage, mRNA-induced oligomerization, and formation of virus-like capsids.

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Series: FEBS journal
ISSN: 1742-464X
ISSN-E: 1742-4658
ISSN-L: 1742-464X
Volume: 288
Issue: 9
Pages: 2930 - 2955
DOI: 10.1111/febs.15618
OADOI: https://oadoi.org/10.1111/febs.15618
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Subjects:
Funding: This work was supported by a Research Council of Norway Toppforsk grant (249951) to CRB, grant PID2019-105872GB-I00/AEI/10.13039/501100011033 (AEI/FEDER, UE) to JMV, and grant MEXT, Japan (20240032, 16H02455, 22110006, 18H04733, and 18H05434), to Y.H. TIRF imaging was performed in the Light Microscopy Facility and Nikon Center of Excellence at the Institute for Applied Life Sciences, University of Massachusetts Amherst, with support from the Massachusetts Life Science Center.
Dataset Reference: S-SAD phasing data: Hallin, Erik I., Touma, Christine, Bramham, Clive R., & Kursula, Petri. (2020). S-SAD dataset for solving the structure of the Arc dimerisation motif [Data set]. Zenodo. http://doi.org/10.5281/zenodo.3765570 ; High-resolution native data: Hallin, Erik I., Touma, Christine, Bramham, Clive R., & Petri Kursula. (2020). High-resolution X-ray diffraction dataset for the coiled-coil oligomerization domain of human Arc [Data set]. Zenodo. http://doi.org/10.5281/zenodo.3766064
  http://doi.org/10.5281/zenodo.3765570
http://doi.org/10.5281/zenodo.3766064
Copyright information: © 2020 Federation of European Biochemical Societies. This is the peer reviewed version of the following article: Eriksen, M.S., Nikolaienko, O., Hallin, E.I., Grødem, S., Bustad, H.J., Flydal, M.I., Merski, I., Hosokawa, T., Lascu, D., Akerkar, S., Cuéllar, J., Chambers, J.J., O’Connell, R., Muruganandam, G., Loris, R., Touma, C., Kanhema, T., Hayashi, Y., Stratton, M.M., Valpuesta, J.M., Kursula, P., Martinez, A. and Bramham, C.R. (2021), Arc self-association and formation of virus-like capsids are mediated by an N-terminal helical coil motif. FEBS J, 288: 2930-2955, which has been published in final form at https://doi.org/10.1111/febs.15618. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.