Mutational analysis of the alpha subunit of eIF2B provides insights into the role of eIF2B bodies in translational control and VWM disease.

NORRIS, Karl, HODGSON, Rachel, DORNELLES, Tawni, ALLEN, K Elizabeth, ABELL, Benjamin, ASHE, Mark P and CAMPBELL, Susan G (2020). Mutational analysis of the alpha subunit of eIF2B provides insights into the role of eIF2B bodies in translational control and VWM disease. Journal of Biological Chemistry.

[img]
Preview
PDF
Norris et al full.pdf - Accepted Version
All rights reserved.

Download (1MB) | Preview
Official URL: https://www.jbc.org/content/early/2020/12/17/jbc.R...
Link to published version:: https://doi.org/10.1074/jbc.RA120.014956
Related URLs:

    Abstract

    Eukaryotic initiation factor 2B (eIF2B) serves as a vital control point within protein synthesis and regulates translation initiation in response to cellular stress. Mutations within eIF2B result in the fatal disease, leukoencephalopathy with vanishing white matter (VWM). Previous biochemical studies on VWM mutations have illustrated that changes in the activity of eIF2B poorly correlates with disease severity. This suggests that there may be additional characteristics of eIF2B contributing to VWM pathogenesis. Here, we investigated whether the localisation of eIF2B to eIF2B bodies was integral for function and whether this localisation could provide insight into the pathogenesis of VWM. We demonstrate that the regulatory subunit, eIF2Bα, is required for the assembly of eIF2B bodies in yeast and that loss of eIF2B bodies correlates with an inability of cells to regulate eIF2B activity.  Mutational analysis of eIF2Bα showed that missense mutations which disrupt the regulation of eIF2B similarly disrupt the assembly of eIF2B bodies. In contrast, when eIF2Bα mutations which impact the catalytic activity of eIF2B were analysed, eIF2B bodies were absent and instead eIF2B localised to small foci, termed microfoci. FRAP analysis highlighted that within these microfoci, eIF2 shuttles more slowly indicating that formation of eIF2B bodies correlates with full eIF2B activity. When eIF2Bα VWM mutations were analysed a diverse impact on localisation was observed, which did not seem to correlate with eIF2B activity.  These findings provide key insights into how the eIF2B body assembles and suggest that the body is a fundamental part of the translational regulation via eIF2α phosphorylation.

    Item Type: Article
    Uncontrolled Keywords: molecular cell biology; protein structure; translation control; translation initiation; translation initiation factor; yeast physiology; Biochemistry & Molecular Biology; 03 Chemical Sciences; 06 Biological Sciences; 11 Medical and Health Sciences
    Identification Number: https://doi.org/10.1074/jbc.RA120.014956
    SWORD Depositor: Symplectic Elements
    Depositing User: Symplectic Elements
    Date Deposited: 22 Dec 2020 12:44
    Last Modified: 22 Dec 2020 12:45
    URI: http://shura.shu.ac.uk/id/eprint/27871

    Actions (login required)

    View Item View Item

    Downloads

    Downloads per month over past year

    View more statistics