title: Structure determination of an 11-subunit exosome in complex with RNA by molecular replacement.
abstract: The RNA exosome is an evolutionarily conserved multi-protein complex involved in the 3' degradation of a variety of RNA transcripts. In the nucleus, the exosome participates in the maturation of structured RNAs, in the surveillance of pre-mRNAs and in the decay of a variety of noncoding transcripts. In the cytoplasm, the exosome degrades mRNAs in constitutive and regulated turnover pathways. Several structures of subcomplexes of eukaryotic exosomes or related prokaryotic exosome-like complexes are known, but how the complete assembly is organized to fulfil processive RNA degradation has been unclear. An atomic snapshot of a Saccharomyces cerevisiae 420 kDa exosome complex bound to an RNA substrate in the pre-cleavage state of a hydrolytic reaction has been determined. Here, the crystallographic steps towards the structural elucidation, which was carried out by molecular replacement, are presented.
authors.affiliation: Structural Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Debora Lika
authors.initials: DL
authors.lastname: Makino
authors.forename: Elena
authors.initials: E
authors.lastname: Conti
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000235:genetics; D019943:Amino Acid Substitution / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D018360:Crystallography, X-Ray; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000235:genetics; D019143:Evolution, Molecular; D063326:Exosome Multienzyme Ribonuclease Complex / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D006868:Hydrolysis; D008958:Models, Molecular; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics; D012322:RNA Precursors / Q000737:chemistry; D012323:RNA Processing, Post-Transcriptional; D062085:RNA, Long Noncoding / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics; D013379:Substrate Specificity / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; D021122:Protein Subunits; D012322:RNA Precursors; D062085:RNA, Long Noncoding; C073751:SRV2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D063326:Exosome Multienzyme Ribonuclease Complex
keywords: RNA; Rrp44; exosome; model building; molecular replacement; nucleases