title: Insights into the structure of the CCR4-NOT complex by electron microscopy.
abstract: The CCR4-NOT complex is a deadenylation complex, which plays a major role for mRNA stability. The complex is conserved from yeast to human and consists of nine proteins NOT1-NOT5, CCR4, CAF1, CAF40 and CAF130. We have successfully isolated the complex using a Protein A tag on NOT1, followed by cross-linking on a glycerol gradient. All components of the complex were identified by mass spectrometry. Electron microscopy of negatively stained particles followed by image reconstruction revealed an L-shaped complex with two arms of similar length. The arms form an accessible cavity, which we think could provide an extensive interface for RNA-deadenylation.
authors.affiliation: European Molecular Biology Laboratory, (EMBL), Heidelberg, Germany. fnasertorabi@ntu.edu.sg
authors.forename: Fariborz
authors.initials: F
authors.lastname: Nasertorabi
authors.forename: Claire
authors.initials: C
authors.lastname: Batisse
authors.forename: Meikel
authors.initials: M
authors.lastname: Diepholz
authors.forename: Dietrich
authors.initials: D
authors.lastname: Suck
authors.forename: Bettina
authors.initials: B
authors.lastname: Böttcher
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000302:isolation & purification*; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012260:Ribonucleases / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure*; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure*
chemical_list: C079592:CDC39 protein, S cerevisiae; D018797:Cell Cycle Proteins; D046912:Multiprotein Complexes; D021122:Protein Subunits; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C078657:CCR4 protein, S cerevisiae; D012260:Ribonucleases; C079009:POP2 protein, S cerevisiae