A Sephin1-insensitive tripartite holophosphatase dephosphorylates translation initiation factor 2α
AffiliationUniv Arizona, Dept Chem & Biochem
Keywordsphosphoprotein phosphatase 1 (PP1)
eukaryotic initiation factor 2 (eIF2)
integrated stress response
MetadataShow full item record
CitationCrespillo-Casado, A., Claes, Z., Choy, M. S., Peti, W., Bollen, M., & Ron, D. (2018). A Sephin1-insensitive tripartite holophosphatase dephosphorylates translation initiation factor 2α. J. Biol. Chem. 2018 293: 7766-. doi:10.1074/jbc.RA118.002325
JournalJOURNAL OF BIOLOGICAL CHEMISTRY
Rights© 2018 Crespillo-Casado et al. Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license.
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AbstractThe integrated stress response (ISR) is regulated by kinases that phosphorylate the subunit of translation initiation factor 2 and phosphatases that dephosphorylate it. Genetic and biochemical observations indicate that the eIF2(P)-directed holophosphatase, a therapeutic target in diseases of protein misfolding, is comprised of a regulatory subunit, PPP1R15, and a catalytic subunit, protein phosphatase 1 (PP1). In mammals, there are two isoforms of the regulatory subunit, PPP1R15A and PPP1R15B, with overlapping roles in the essential function of eIF2(P) dephosphorylation. However, conflicting reports have appeared regarding the requirement for an additional co-factor, G-actin, in enabling substrate-specific dephosphorylation by PPP1R15-containing PP1 holoenzymes. An additional concern relates to the sensitivity of the holoenzyme to the [(o-chlorobenzylidene)amino]guanidines Sephin1 or guanabenz, putative small-molecule proteostasis modulators. It has been suggested that the source and method of purification of the PP1 catalytic subunit and the presence or absence of an N-terminal repeat-containing region in the PPP1R15A regulatory subunit might influence the requirement for G-actin and sensitivity of the holoenzyme to inhibitors. We found that eIF2(P) dephosphorylation by PP1 was moderately stimulated by repeat-containing PPP1R15A in an unphysiological low ionic strength buffer, whereas stimulation imparted by the co-presence of PPP1R15A and G-actin was observed under a broad range of conditions, low and physiological ionic strength, regardless of whether the PPP1R15A regulatory subunit had or lacked the N-terminal repeat-containing region and whether it was paired with native PP1 purified from rabbit muscle or recombinant PP1 purified from bacteria. Furthermore, none of the PPP1R15A-containing holophosphatases tested were inhibited by Sephin1 or guanabenz.
Note12 month embargo; published online: 4 April 2018
VersionFinal published version
SponsorsWellcome Trust principal research fellowship [Wellcome 200848/Z/16/Z]; Wellcome Trust strategic award [Wellcome 100140]; National Institute of Health [R01NS091336]; American Diabetes Association Pathway to Stop Diabetes [1-14-ACN-31]; Flemish Concerted Research Action [GOA15/016]
- PPP1R15A-mediated dephosphorylation of eIF2α is unaffected by Sephin1 or Guanabenz.
- Authors: Crespillo-Casado A, Chambers JE, Fischer PM, Marciniak SJ, Ron D
- Issue date: 2017 Apr 27
- Actin dynamics tune the integrated stress response by regulating eukaryotic initiation factor 2α dephosphorylation.
- Authors: Chambers JE, Dalton LE, Clarke HJ, Malzer E, Dominicus CS, Patel V, Moorhead G, Ron D, Marciniak SJ
- Issue date: 2015 Mar 16
- Decoding the selectivity of eIF2α holophosphatases and PPP1R15A inhibitors.
- Authors: Carrara M, Sigurdardottir A, Bertolotti A
- Issue date: 2017 Sep
- G-actin provides substrate-specificity to eukaryotic initiation factor 2α holophosphatases.
- Authors: Chen R, Rato C, Yan Y, Crespillo-Casado A, Clarke HJ, Harding HP, Marciniak SJ, Read RJ, Ron D
- Issue date: 2015 Mar 16
- Selective inhibition of a regulatory subunit of protein phosphatase 1 restores proteostasis.
- Authors: Tsaytler P, Harding HP, Ron D, Bertolotti A
- Issue date: 2011 Apr 1