title: Large terminase conformational change induced by connector binding in bacteriophage T7.
abstract: During bacteriophage morphogenesis DNA is translocated into a preformed prohead by the complex formed by the portal protein, or connector, plus the terminase, which are located at an especial prohead vertex. The terminase is a powerful motor that converts ATP hydrolysis into mechanical movement of the DNA. Here, we have determined the structure of the T7 large terminase by electron microscopy. The five terminase subunits assemble in a toroid that encloses a channel wide enough to accommodate dsDNA. The structure of the complete connector-terminase complex is also reported, revealing the coupling between the terminase and the connector forming a continuous channel. The structure of the terminase assembled into the complex showed a different conformation when compared with the isolated terminase pentamer. To understand in molecular terms the terminase morphological change, we generated the terminase atomic model based on the crystallographic structure of its phage T4 counterpart. The docking of the threaded model in both terminase conformations showed that the transition between the two states can be achieved by rigid body subunit rotation in the pentameric assembly. The existence of two terminase conformations and its possible relation to the sequential DNA translocation may shed light into the molecular bases of the packaging mechanism of bacteriophage T7.
authors: Department of Macromolecular Structure, 28049 Madrid, Spain.
authors: María I
authors: MI
authors: Daudén
authors: Department of Macromolecular Structure, 28049 Madrid, Spain.
authors: Jaime
authors: J
authors: Martín-Benito
authors: Computational Systems Biology Group, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, 28049 Madrid, Spain.
authors: Juan C
authors: JC
authors: Sánchez-Ferrero
authors: Department of Macromolecular Structure, 28049 Madrid, Spain.
authors: Mar
authors: M
authors: Pulido-Cid
authors: Department of Macromolecular Structure, 28049 Madrid, Spain.
authors: José M
authors: JM
authors: Valpuesta
authors: Department of Macromolecular Structure, 28049 Madrid, Spain. Electronic address: jlcarras@cnb.csic.es.
authors: José L
authors: JL
authors: Carrascosa
mesh_terms: D017123:Bacteriophage T7 / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure; D004279:DNA, Viral / Q000737:chemistry* / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology; D062105:Molecular Docking Simulation; D020836:Protein Structure, Quaternary; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism; D019065:Virus Assembly / Q000502:physiology
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins; D004706:Endodeoxyribonucleases; C030894:terminase
keywords: ATPases; Bacteriophage; DNA Packaging; DNA Translocation; Electron Microscopy (EM); Protein Conformation; Single Particle Reconstruction; Terminase; Virus Assembly
references: J Struct Biol. 1996 Oct;116(1):237-40
references: 8812978
references: J Biol Chem. 1982 Nov 25;257(22):13223-32
references: 6815174
references: J Mol Biol. 2007 May 25;369(1):79-94
references: 17428497
references: J Mol Biol. 1994 Jan 7;235(1):248-59
references: 8289246
references: EMBO J. 2003 Mar 17;22(6):1255-62
references: 12628918
references: Virology. 1986 May;151(1):119-23
references: 3754362
references: J Mol Biol. 1971 Dec 14;62(2):287-301
references: 4944929
references: Cell. 2005 Sep 9;122(5):683-92
references: 16143101
references: Virology. 1991 Dec;185(2):788-94
references: 1962450
references: Cell. 2004 Sep 17;118(6):743-55
references: 15369673
references: J Mol Biol. 2003 Aug 1;331(1):139-54
references: 12875841
references: J Biol Chem. 2000 Nov 24;275(47):37127-36
references: 10967092
references: Curr Opin Biotechnol. 2012 Aug;23(4):529-36
references: 22186221
references: Curr Opin Struct Biol. 2005 Dec;15(6):655-63
references: 16271469
references: J Mol Biol. 1987 Sep 20;197(2):229-36
references: 2960820
references: J Struct Biol. 2000 Aug;131(2):146-55
references: 11042085
references: Nat Methods. 2008 Jan;5(1):53-5
references: 18157137
references: J Biol Chem. 2006 Aug 4;281(31):21914-21923
references: 16735502
references: Nat Methods. 2007 Jan;4(1):27-9
references: 17179934
references: J Struct Biol. 2004 Nov;148(2):194-204
references: 15477099
references: J Struct Biol. 2010 Aug;171(2):197-206
references: 20362059
references: Proc Natl Acad Sci U S A. 2007 Oct 23;104(43):16868-73
references: 17942694
references: J Struct Biol. 1999 Dec 1;128(1):82-97
references: 10600563
references: Nat Rev Mol Cell Biol. 2005 Jul;6(7):519-29
references: 16072036
references: J Comput Chem. 2004 Oct;25(13):1605-12
references: 15264254
references: Structure. 2007 Apr;15(4):461-72
references: 17437718
references: Proc Natl Acad Sci U S A. 1978 Oct;75(10):4779-83
references: 283391
references: J Virol. 2012 Apr;86(8):4046-57
references: 22345478
references: Electrophoresis. 1997 Dec;18(15):2714-23
references: 9504803
references: J Mol Biol. 2008 Nov 14;383(3):494-501
references: 18775728
references: Nature. 2000 Dec 7;408(6813):745-50
references: 11130079
references: J Mol Biol. 1987 Aug 20;196(4):845-51
references: 3316664
references: Biochim Biophys Acta. 2000 May 31;1458(2-3):482-510
references: 10838060
references: Nucleic Acids Res. 2005 Jul 1;33(Web Server issue):W244-8
references: 15980461
references: Nanotechnology. 2012 Jan 13;23(1):015501
references: 22156040
references: Structure. 2012 Aug 8;20(8):1403-13
references: 22771211
references: J Biol Chem. 2011 Jan 7;286(1):234-42
references: 20962334
references: Nature. 2009 Jan 22;457(7228):446-50
references: 19129763
references: Nat Struct Mol Biol. 2004 Feb;11(2):142-8
references: 14730353
references: Bioinformatics. 2005 Sep 1;21 Suppl 2:ii243-4
references: 16204112
references: J Struct Biol. 1999 Apr-May;125(2-3):185-95
references: 10222274
references: J Mol Biol. 2007 Nov 9;373(5):1113-22
references: 17919653
references: J Mol Biol. 2002 Jan 25;315(4):663-76
references: 11812138
references: Acta Crystallogr D Biol Crystallogr. 1999 Sep;55(Pt 9):1495-502
references: 10489444
references: Nature. 1998 Nov 19;396(6708):279-82
references: 9834036
references: EMBO J. 2005 Nov 2;24(21):3820-9
references: 16211007
references: Nature. 2009 Oct 1;461(7264):669-73
references: 19794496
references: Virology. 1991 Feb;180(2):709-15
references: 1989388
references: J Mol Biol. 1996 Aug 30;261(4):524-35
references: 8794874
references: Virology. 1986 May;151(1):110-8
references: 3962187
references: Cell. 2008 Dec 26;135(7):1251-62
references: 19109896
references: J Mol Biol. 2007 Nov 30;374(3):817-36
references: 17945256
references: Mol Cell. 2002 May;9(5):981-91
references: 12049735
references: Adv Protein Chem. 2003;64:301-23
references: 13677051
references: Virology. 1987 Aug;159(2):250-8
references: 3617498
references: J Biol Chem. 2012 Aug 10;287(33):28196-205
references: 22715098
references: Biol Proced Online. 2004;6:23-34
references: 15103397
references: J Mol Biol. 2005 Apr 15;347(5):895-902
references: 15784250
references: J Virol. 2004 Nov;78(22):12668-71
references: 15507654
references: PLoS Biol. 2007 Mar;5(3):e59
references: 17311473
references: Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16078-83
references: 20805464
references: Mol Cell. 2007 Mar 23;25(6):943-9
references: 17386269
references: J Biol Chem. 2000 Nov 10;275(45):35311-9
references: 10930407
references: Virology. 1987 Aug;159(2):244-9
references: 2956757
references: Nat Struct Biol. 1999 Sep;6(9):842-6
references: 10467096
references: Virology. 1993 Apr;193(2):748-52
references: 8460483
references: Mol Cell. 2006 Aug;23(4):457-69
references: 16916635
references: Mol Microbiol. 2006 Jul;61(1):16-32
references: 16824092
references: Proc Natl Acad Sci U S A. 2012 Jan 17;109(3):817-22
references: 22207623
references: Nature. 2001 Oct 18;413(6857):748-52
references: 11607035
references: Nat Rev Microbiol. 2011 Aug 12;9(9):647-57
references: 21836625
references: J Struct Biol. 1996 Jan-Feb;116(1):190-9
references: 8742743
references: J Mol Biol. 2012 Oct 26;423(3):413-26
references: 22858866
references: Virology. 1995 Aug 20;211(2):516-24
references: 7645255
references: Curr Opin Microbiol. 2006 Aug;9(4):423-9
references: 16814597
references: J Mol Biol. 1995 Feb 3;245(5):635-44
references: 7844832
references: Cell. 2000 Jun 9;101(6):589-600
references: 10892646
references: Annu Rev Genet. 2008;42:647-81
references: 18687036
references: Q Rev Biophys. 1994 May;27(2):107-155
references: 7984775
references: EMBO Rep. 2009 Jun;10(6):592-8
references: 19444313
references: Proc Natl Acad Sci U S A. 2012 Jan 17;109(3):811-6
references: 22207627
references: Biophys J. 2008 Jan 1;94(1):159-67
references: 17827233
references: Cell. 2006 Dec 29;127(7):1349-60
references: 17190599
references: J Struct Biol. 2008 Oct;164(1):170-5
references: 18614378