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

Maria Eriksen, Oleksii Nikolaienko, Erik Hallin, Sverre Grødem, Helene Bustad, Marte Flydal, Ian Merski, Tomohisa Hosokawa, Daniela Lascu, Shreeram Akerkar, Jorge Cuéllar, James Chambers, Rory O'Connell, Gopinath Muruganandam, Remy Loris, Christine Touma, Tambudzai Kanhema, Yasunori Hayashi, Margaret Stratton, Jose ValpuestaPetri Kursula, Aurora Martinez, Clive Bramham

Onderzoeksoutput: Articlepeer review

14 Citaten (Scopus)
27 Downloads (Pure)

Samenvatting

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. Database: The coordinates and structure factors for crystallographic analysis of the oligomerization region were deposited at the Protein Data Bank with the entry code 6YTU.

Originele taal-2English
Pagina's (van-tot)2930-2955
Aantal pagina's26
TijdschriftThe FEBS Journal
Volume288
Nummer van het tijdschrift9
Vroegere onlinedatum11 nov 2020
DOI's
StatusPublished - 3 mei 2021

Bibliografische nota

Funding Information:
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. We gratefully acknowledge beamtime and beamline support at Diamond Light Source, EMBL/DESY, and SOLEIL.

Funding Information:
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. We gratefully acknowledge beamtime and beamline support at Diamond Light Source, EMBL/DESY, and SOLEIL.

Publisher Copyright:
© 2020 Federation of European Biochemical Societies

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Vingerafdruk

Duik in de onderzoeksthema's van 'Arc self‐association and formation of virus‐like capsids are mediated by an N‐terminal helical coil motif'. Samen vormen ze een unieke vingerafdruk.

Citeer dit