title: Direct interaction between EFL1 and SBDS is mediated by an intrinsically disordered insertion domain.
abstract: Removal of anti-association factor, Tif6 (eIF6), by elongation factor-like 1 (EFL1) and Shwachman-Bodian-Diamond syndrome (SBDS) protein is a critical step in the late stage of ribosome maturation. Although EFL1 is known to have GTPase activity that is stimulated by SBDS, how they cooperatively trigger dissociation of Tif6 from the ribosome remains to be elucidated. In the present study, the interaction between EFL1 and SBDS was analyzed by size exclusion chromatography, gel shift assay, and isothermal titration calorimetry (ITC). The results showed that EFL1 interacted directly with SBDS. ITC experiments using domain-truncated mutants showed that the interaction between EFL1 and SBDS is governed by the insertion domain of EFL1 and domains II-III of SBDS. Circular dichroism spectroscopy showed that the insertion domain of EFL1 has a random structure in the absence of SBDS, whereas the disadvantageous entropy change observed on ITC suggested a fixed conformation coupled with complex formation with SBDS. Based on these observations together with those reported previously, we propose roles of EFL1 and SBDS in ribosomal maturation.
authors.affiliation: Graduate School of Life Sciences, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Nozomi
authors.initials: N
authors.lastname: Asano
authors.affiliation: Graduate School of Life Sciences, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Haruka
authors.initials: H
authors.lastname: Atsuumi
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Akiyoshi
authors.initials: A
authors.lastname: Nakamura
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Tanaka
authors.affiliation: Faculty of Advanced Life Science, Hokkaido University, Sapporo 060-0810, Japan. Electronic address: yao@castor.sci.hokudai.ac.jp.
authors.forename: Min
authors.initials: M
authors.lastname: Yao
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D001855:Bone Marrow Diseases / Q000235:genetics / Q000378:metabolism; D010188:Exocrine Pancreatic Insufficiency / Q000235:genetics / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D064267:Intrinsically Disordered Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008068:Lipomatosis / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D020652:Peptide Elongation Factor 2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D000081003:Shwachman-Diamond Syndrome; D013816:Thermodynamics
chemical_list: D064267:Intrinsically Disordered Proteins; D020652:Peptide Elongation Factor 2; D011506:Proteins; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C470521:SBDS protein, human; D029701:Saccharomyces cerevisiae Proteins; C491609:TIF6 protein, S cerevisiae; D020558:GTP Phosphohydrolases; C520003:RIA1 protein, S cerevisiae
keywords: Circular dichroism; Interaction; Intrinsically disordered protein; Isothermal titration calorimetry; Ribosome biogenesis