title: Transposition into replicating DNA occurs through interaction with the processivity factor.
abstract: The bacterial transposon Tn7 directs transposition into actively replicating DNA by a mechanism involving the transposon-encoded protein TnsE. Here we show that TnsE physically and functionally interacts with the processivity factor of the DNA replication machinery in vivo and in vitro. Our work establishes an in vitro TnsABC+E transposition reaction reconstituted from purified proteins and target DNA structures. Using the in vitro reaction we confirm that the processivity factor specifically reorders TnsE-mediated transposition events on target DNAs in a way that matches the bias with active DNA replication in vivo. The TnsE interaction with an essential and conserved component of the replication machinery, and a DNA structure reveals a mechanism by which Tn7, and probably other elements, selects target sites associated with DNA replication.
authors.affiliation: Department of Microbiology, Cornell University, Ithaca, NY 14853, USA.
authors.forename: Adam R
authors.initials: AR
authors.lastname: Parks
authors.forename: Zaoping
authors.initials: Z
authors.lastname: Li
authors.forename: Qiaojuan
authors.initials: Q
authors.lastname: Shi
authors.forename: Roisin M
authors.initials: RM
authors.lastname: Owens
authors.forename: Moonsoo M
authors.initials: MM
authors.lastname: Jin
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Peters
mesh_terms: D000595:Amino Acid Sequence; D004258:DNA Polymerase III / Q000378:metabolism; D004261:DNA Replication; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016415:Sequence Alignment
chemical_list: D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C424904:TnsE protein, E coli; C101139:beta subunit, DNA polymerase III; D004258:DNA Polymerase III
references.citation: Genetics. 2005 Oct;171(2):503-15
references.pmid: 16085704
references.citation: Nucleic Acids Res. 2006 Apr 28;34(8):e57
references.pmid: 16648358
references.citation: EMBO J. 2003 Nov 3;22(21):5883-92
references.pmid: 14592985
references.citation: Genetics. 1997 Mar;145(3):573-85
references.pmid: 9055068
references.citation: Bioinformatics. 2004 Feb 12;20(3):426-7
references.pmid: 14960472
references.citation: Genetics. 2008 Jul;179(3):1237-50
references.pmid: 18562643
references.citation: J Bacteriol. 2004 Nov;186(21):7280-9
references.pmid: 15489440
references.citation: Philos Trans R Soc Lond B Biol Sci. 2004 Jan 29;359(1441):25-30
references.pmid: 15065653
references.citation: J Mol Biol. 2001 Jan 19;305(3):633-42
references.pmid: 11152618
references.citation: Methods Enzymol. 1995;262:430-42
references.pmid: 8594367
references.citation: Mol Biotechnol. 2001 Oct;19(2):133-40
references.pmid: 11725483
references.citation: Cell. 2001 Aug 24;106(4):417-28
references.pmid: 11525728
references.citation: J Mol Biol. 2001 Apr 13;307(5):1161-70
references.pmid: 11292332
references.citation: J Biol Chem. 2008 Aug 22;283(34):22895-906
references.pmid: 18556658
references.citation: CSH Protoc. 2006 Jun 01;2006(1):
references.pmid: 22485559
references.citation: Cell. 2001 Aug 24;106(4):429-41
references.pmid: 11525729
references.citation: Genes Dev. 2001 Mar 15;15(6):737-47
references.pmid: 11274058
references.citation: Mol Microbiol. 2007 Aug;65(3):811-27
references.pmid: 17635192
references.citation: Nucleic Acids Res. 1998 Sep 1;26(17):3925-32
references.pmid: 9705499
references.citation: J Bacteriol. 2004 Oct;186(20):6738-48
references.pmid: 15466025
references.citation: Genes Dev. 1996 Sep 1;10(17):2145-57
references.pmid: 8804309
references.citation: CSH Protoc. 2007 Aug 01;2007:pdb.prot4759
references.pmid: 21357139
references.citation: Nat Rev Mol Cell Biol. 2001 Nov;2(11):806-14
references.pmid: 11715047
references.citation: Trends Biochem Sci. 1998 Dec;23(12):486-90
references.pmid: 9868372
references.citation: J Bacteriol. 1988 Jan;170(1):352-8
references.pmid: 2826397
references.citation: Cell. 2008 Jan 11;132(1):43-54
references.pmid: 18191219
references.citation: Plasmid. 2009 Jan;61(1):1-14
references.pmid: 18951916
references.citation: Biochemistry. 2004 May 18;43(19):5661-71
references.pmid: 15134440
references.citation: Genetics. 1992 Mar;130(3):665-76
references.pmid: 1312981
references.citation: J Bacteriol. 2007 Mar;189(5):2170-3
references.pmid: 17194796
references.citation: J Biol Chem. 2006 May 19;281(20):14340-9
references.pmid: 16546997
references.citation: Cell. 1993 Mar 26;72(6):931-43
references.pmid: 8384534
references.citation: Nature. 1989 Jul 20;340(6230):245-6
references.pmid: 2547163
references.citation: Appl Environ Microbiol. 2008 Jun;74(11):3419-25
references.pmid: 18408063
references.citation: Bioessays. 1995 Aug;17(8):733-41
references.pmid: 7661854
references.citation: J Biol Chem. 2000 Nov 3;275(44):34609-18
references.pmid: 10924523
references.citation: J Mol Biol. 1982 Oct 15;161(1):57-76
references.pmid: 6818353
references.citation: Mol Cell. 2008 Feb 15;29(3):291-301
references.pmid: 18280235
references.citation: Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8376-80
references.pmid: 11459978
references.citation: EMBO J. 2001 Feb 15;20(4):924-32
references.pmid: 11179236
references.citation: Nucleic Acids Res. 2000 Mar 1;28(5):1067-77
references.pmid: 10666445
references.citation: Gene. 1995 Dec 1;166(1):177-8
references.pmid: 8529886
references.citation: Mol Microbiol. 2005 Sep;57(6):1664-75
references.pmid: 16135232
references.citation: Nat Rev Mol Cell Biol. 2006 May;7(5):335-46
references.pmid: 16612326
references.citation: Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11627-32
references.pmid: 11573000
references.citation: Annu Rev Biochem. 2005;74:283-315
references.pmid: 15952889
references.citation: EMBO J. 1998 Oct 1;17(19):5822-31
references.pmid: 9755182
references.citation: Bioessays. 2000 Nov;22(11):997-1006
references.pmid: 11056476
references.citation: Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3936-41
references.pmid: 10737770
references.citation: J Biol Chem. 1995 Dec 8;270(49):29563-9
references.pmid: 7493999
references.citation: J Biol Chem. 2001 Dec 14;276(50):47185-94
references.pmid: 11572866
references.citation: Mol Cell. 2000 Sep;6(3):573-82
references.pmid: 11030337
references.citation: Mol Microbiol. 2006 Jan;59(2):460-74
references.pmid: 16390442
references.citation: Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
references.pmid: 7984417