G-actin provides substrate-specificity to eukaryotic initiation factor 2α holophosphatases.

Abstract:

:Dephosphorylation of eukaryotic translation initiation factor 2a (eIF2a) restores protein synthesis at the waning of stress responses and requires a PP1 catalytic subunit and a regulatory subunit, PPP1R15A/GADD34 or PPP1R15B/CReP. Surprisingly, PPP1R15-PP1 binary complexes reconstituted in vitro lacked substrate selectivity. However, selectivity was restored by crude cell lysate or purified G-actin, which joined PPP1R15-PP1 to form a stable ternary complex. In crystal structures of the non-selective PPP1R15B-PP1G complex, the functional core of PPP1R15 made multiple surface contacts with PP1G, but at a distance from the active site, whereas in the substrate-selective ternary complex, actin contributes to one face of a platform encompassing the active site. Computational docking of the N-terminal lobe of eIF2a at this platform placed phosphorylated serine 51 near the active site. Mutagenesis of predicted surface-contacting residues enfeebled dephosphorylation, suggesting that avidity for the substrate plays an important role in imparting specificity on the PPP1R15B-PP1G-actin ternary complex.

journal_name

Elife

journal_title

eLife

authors

Chen R,Rato C,Yan Y,Crespillo-Casado A,Clarke HJ,Harding HP,Marciniak SJ,Read RJ,Ron D

doi

10.7554/eLife.04871

subject

Has Abstract

pub_date

2015-03-16 00:00:00

issn

2050-084X

journal_volume

4

pub_type

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