title: Conserved beta-hairpin recognition by the GYF domains of Smy2 and GIGYF2 in mRNA surveillance and vesicular transport complexes.
abstract: The yeast suppressor of myosin 2 protein (Smy2) interacts with mRNA-processing proteins through recognition of proline-rich sequences (PRS). Here, we describe the crystal structure of the GYF domain of Smy2 in association with a PRS from the yeast branch point binding protein (BBP/ScSF1). Complex formation requires that the beta-hairpin of the central PPGL motif of the ligand is accommodated by an extended hydrophobic cleft in the domain-a specificity feature that is maintained in the human protein GIGYF2. SILAC/MS experiments in combination with PRS site inhibition show that Smy2 associates with the Ccr4-NOT deadenylase complex, whereas GIGYF2 interacts not only with mRNA surveillance factors, but also with vesicular transport proteins and Atrophin-1. GIGYF2 is shown to associate with COPII-vesicle proteins and localize to the ER and Golgi in resting cells, whereas environmental challenge drives GIGYF2 into stress granules. The current study highlights the structural basis for PRS recognition by Smy2-type GYF domains, and implicates Smy2 and GIGYF2 in both mRNA processing and the secretory pathway.
authors.affiliation: Protein Engineering, Leibniz-Institut fuer Molekulare Pharmakologie, 13125 Berlin, Germany.
authors.forename: Miriam-Rose
authors.initials: MR
authors.lastname: Ash
authors.forename: Katja
authors.initials: K
authors.lastname: Faelber
authors.forename: Daniela
authors.initials: D
authors.lastname: Kosslick
authors.forename: Gesa Ines
authors.initials: GI
authors.lastname: Albert
authors.forename: Yvette
authors.initials: Y
authors.lastname: Roske
authors.forename: Michael
authors.initials: M
authors.lastname: Kofler
authors.forename: Michael
authors.initials: M
authors.lastname: Schuemann
authors.forename: Eberhard
authors.initials: E
authors.lastname: Krause
authors.forename: Christian
authors.initials: C
authors.lastname: Freund
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D003460:Crystallization; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional / Q000502:physiology*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics; D017422:Sequence Analysis, DNA; D022161:Transport Vesicles / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D002352:Carrier Proteins; C478016:GIGYF2 protein, human; C083574:SMY2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins