title: The translation initiation factor eIF2beta is an interactor of protein phosphatase-1.
abstract: It is reasonably well understood how the initiation of translation is controlled by reversible phosphorylation of the eukaryotic translation initiation factors eIF2alpha, eIF2Bepsilon and eIF4E. Other initiation factors, including eIF2beta, are also established phosphoproteins but the physiological impact of their phosphorylation is not known. Using a sequence homology search we found that the central region of eIF2beta contains a putative PP1-(protein phosphatase-1) binding RVxF-motif. The predicted eIF2beta-PP1 interaction was confirmed by PP1 binding and co-immunoprecipitation assays on cell lysates as well as with the purified components. Site-directed mutagenesis showed that eIF2beta contains, in addition to an RVxF-motif, at least one other PP1-binding site in its C-terminal half. eIF2beta functioned as an inhibitor for the dephosphorylation of glycogen phosphorylase and Ser51 of eIF2alpha by PP1, but did not affect the dephosphorylation of Ser464 of eIF2Bepsilon by this phosphatase. Strikingly, eIF2beta emerged as an activator of its own dephosphorylation (Ser2, Ser67, Ser218) by associated PP1, since the substrate quality of eIF2beta was decreased by the mere mutation of its RVxF-motif. These results make eIF2beta an attractive candidate substrate for associated PP1 in vivo. The overexpression of wild-type eIF2beta or eIF2beta with a mutated RVxF-motif did not differentially affect the rate of translation, indicating that the binding of PP1 is not rate-limiting for translation under basal conditions.
authors.affiliation: Department of Molecular Cell Biology, Division of Biochemistry, Faculty of Medicine, Katholieke Universiteit Leuven, B3000 Leuven, Belgium.
authors.forename: Paulina
authors.initials: P
authors.lastname: Wakula
authors.forename: Monique
authors.initials: M
authors.lastname: Beullens
authors.forename: Aleyde
authors.initials: A
authors.lastname: van Eynde
authors.forename: Hugo
authors.initials: H
authors.lastname: Ceulemans
authors.forename: Willy
authors.initials: W
authors.lastname: Stalmans
authors.forename: Mathieu
authors.initials: M
authors.lastname: Bollen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D014176:Protein Biosynthesis; D054645:Protein Phosphatase 1; D011817:Rabbits; D016415:Sequence Alignment; D013379:Substrate Specificity; D013552:Swine
chemical_list: D015852:Eukaryotic Initiation Factor-2; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1