title: Structure of EF-G-ribosome complex in a pretranslocation state.
abstract: In protein synthesis, elongation factor G (EF-G) facilitates movement of tRNA-mRNA by one codon, which is coupled to the ratchet-like rotation of the ribosome complex and is triggered by EF-G-mediated GTP hydrolysis. Here we report the structure of a pretranslocational ribosome bound to Thermus thermophilus EF-G trapped with a GTP analog. The positioning of the catalytic His87 into the active site coupled to hydrophobic-gate opening involves the 23S rRNA sarcin-ricin loop and domain III of EF-G and provides a structural basis for the GTPase activation of EF-G. Interactions of the hybrid peptidyl-site-exit-site tRNA with ribosomal elements, including the entire L1 stalk and proteins S13 and S19, shed light on how formation and stabilization of the hybrid tRNA is coupled to head swiveling and body rotation of the 30S as well as to closure of the L1 stalk.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore.
authors.forename: Yun
authors.initials: Y
authors.lastname: Chen
authors.forename: Shu
authors.initials: S
authors.lastname: Feng
authors.forename: Veerendra
authors.initials: V
authors.lastname: Kumar
authors.forename: Rya
authors.initials: R
authors.lastname: Ero
authors.forename: Yong-Gui
authors.initials: YG
authors.lastname: Gao
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006160:Guanosine Triphosphate / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020653:Peptide Elongation Factor G / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D016963:Thermus thermophilus / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D020653:Peptide Elongation Factor G; D012329:RNA, Bacterial; D012333:RNA, Messenger; D006160:Guanosine Triphosphate; D012343:RNA, Transfer
references.citation: Science. 2011 Jul 1;333(6038):37; author reply 37
references.pmid: 21719661
references.citation: Mol Cell. 2003 Aug;12(2):321-8
references.pmid: 14536072
references.citation: Biochimie. 1987 Sep;69(9):911-23
references.pmid: 3126829
references.citation: Nat Commun. 2013;4:1733
references.pmid: 23591900
references.citation: EMBO J. 1998 Jun 15;17(12):3478-83
references.pmid: 9628883
references.citation: Proc Natl Acad Sci U S A. 2011 Sep 20;108(38):15798-803
references.pmid: 21903932
references.citation: Cell. 2003 Jul 11;114(1):123-34
references.pmid: 12859903
references.citation: Nat Struct Mol Biol. 2006 Mar;13(3):234-41
references.pmid: 16501572
references.citation: Science. 2011 Dec 16;334(6062):1524-9
references.pmid: 22096102
references.citation: Science. 2013 Jun 28;340(6140):1235970
references.pmid: 23812721
references.citation: Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
references.pmid: 15299374
references.citation: Proc Natl Acad Sci U S A. 2009 Jan 27;106(4):1063-8
references.pmid: 19122150
references.citation: Mol Cell. 2007 Mar 9;25(5):751-64
references.pmid: 17349960
references.citation: J Mol Biol. 2008 Sep 26;382(1):179-87
references.pmid: 18644383
references.citation: J Mol Biol. 2005 May 13;348(4):939-49
references.pmid: 15843024
references.citation: Mol Cell. 2008 Oct 24;32(2):190-7
references.pmid: 18951087
references.citation: Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
references.pmid: 9757107
references.citation: J Biol Chem. 2000 Nov 17;275(46):35820-4
references.pmid: 10940297
references.citation: J Mol Biol. 2010 Oct 1;402(4):741-60
references.pmid: 20691699
references.citation: Nat Chem Biol. 2010 May;6(5):344-51
references.pmid: 20348921
references.citation: EMBO J. 2007 May 2;26(9):2421-31
references.pmid: 17446867
references.citation: Nat Struct Biol. 1995 Jan;2(1):36-44
references.pmid: 7719852
references.citation: Science. 2011 May 20;332(6032):981-4
references.pmid: 21596992
references.citation: EMBO J. 2009 Mar 18;28(6):755-65
references.pmid: 19229291
references.citation: Biol Chem. 2000 May-Jun;381(5-6):367-75
references.pmid: 10937867
references.citation: Nature. 1989 Nov 9;342(6246):142-8
references.pmid: 2682263
references.citation: Nat Struct Mol Biol. 2010 Sep;17(9):1072-8
references.pmid: 20694005
references.citation: EMBO J. 1994 Aug 15;13(16):3661-8
references.pmid: 8070396
references.citation: J Mol Biol. 2005 May 27;349(1):47-59
references.pmid: 15876367
references.citation: EMBO J. 1994 Aug 15;13(16):3669-77
references.pmid: 8070397
references.citation: RNA. 2012 Feb;18(2):230-40
references.pmid: 22187675
references.citation: Biochem J. 1989 Apr 1;259(1):307-10
references.pmid: 2655578
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references.pmid: 15572765
references.citation: PLoS One. 2013;8(3):e58829
references.pmid: 23527033
references.citation: Nat Struct Mol Biol. 2006 Apr;13(4):354-9
references.pmid: 16532005
references.citation: Science. 2013 Jun 28;340(6140):1235490
references.pmid: 23812720
references.citation: Science. 2009 Oct 30;326(5953):694-9
references.pmid: 19833919
references.citation: Acta Crystallogr D Biol Crystallogr. 2012 May;68(Pt 5):578-83
references.pmid: 22525755
references.citation: Mol Cell. 2003 Jun;11(6):1517-23
references.pmid: 12820965
references.citation: Science. 2013 Jun 28;340(6140):1236086
references.pmid: 23812722
references.citation: Proc Natl Acad Sci U S A. 2009 Sep 15;106(37):15702-7
references.pmid: 19717422
references.citation: Nature. 2010 Dec 2;468(7324):713-6
references.pmid: 21124459
references.citation: EMBO J. 2005 Dec 21;24(24):4316-23
references.pmid: 16292341
references.citation: J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
references.pmid: 19461840
references.citation: Mol Cell. 2007 Feb 23;25(4):505-17
references.pmid: 17317624
references.citation: Nature. 2000 Jul 20;406(6793):318-22
references.pmid: 10917535
references.citation: EMBO J. 1989 Dec 1;8(12):3933-8
references.pmid: 2583120
references.citation: Science. 2010 Nov 5;330(6005):835-838
references.pmid: 21051640
references.citation: Nature. 1997 Jan 2;385(6611):37-41
references.pmid: 8985244