Transposition into replicating DNA occurs through interaction with the processivity factor.
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.
Parks
Adam R
AR
Department of Microbiology, Cornell University, Ithaca, NY 14853, USA.
Li
Zaoping
Z
Shi
Qiaojuan
Q
Owens
Roisin M
RM
Jin
Moonsoo M
MM
Peters
Joseph E
JE
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
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
Genetics. 2005 Oct;171(2):503-15
16085704
Nucleic Acids Res. 2006 Apr 28;34(8):e57
16648358
EMBO J. 2003 Nov 3;22(21):5883-92
14592985
Genetics. 1997 Mar;145(3):573-85
9055068
Bioinformatics. 2004 Feb 12;20(3):426-7
14960472
Genetics. 2008 Jul;179(3):1237-50
18562643
J Bacteriol. 2004 Nov;186(21):7280-9
15489440
Philos Trans R Soc Lond B Biol Sci. 2004 Jan 29;359(1441):25-30
15065653
J Mol Biol. 2001 Jan 19;305(3):633-42
11152618
Methods Enzymol. 1995;262:430-42
8594367
Mol Biotechnol. 2001 Oct;19(2):133-40
11725483
Cell. 2001 Aug 24;106(4):417-28
11525728
J Mol Biol. 2001 Apr 13;307(5):1161-70
11292332
J Biol Chem. 2008 Aug 22;283(34):22895-906
18556658
CSH Protoc. 2006 Jun 01;2006(1):
22485559
Cell. 2001 Aug 24;106(4):429-41
11525729
Genes Dev. 2001 Mar 15;15(6):737-47
11274058
Mol Microbiol. 2007 Aug;65(3):811-27
17635192
Nucleic Acids Res. 1998 Sep 1;26(17):3925-32
9705499
J Bacteriol. 2004 Oct;186(20):6738-48
15466025
Genes Dev. 1996 Sep 1;10(17):2145-57
8804309
CSH Protoc. 2007 Aug 01;2007:pdb.prot4759
21357139
Nat Rev Mol Cell Biol. 2001 Nov;2(11):806-14
11715047
Trends Biochem Sci. 1998 Dec;23(12):486-90
9868372
J Bacteriol. 1988 Jan;170(1):352-8
2826397
Cell. 2008 Jan 11;132(1):43-54
18191219
Plasmid. 2009 Jan;61(1):1-14
18951916
Biochemistry. 2004 May 18;43(19):5661-71
15134440
Genetics. 1992 Mar;130(3):665-76
1312981
J Bacteriol. 2007 Mar;189(5):2170-3
17194796
J Biol Chem. 2006 May 19;281(20):14340-9
16546997
Cell. 1993 Mar 26;72(6):931-43
8384534
Nature. 1989 Jul 20;340(6230):245-6
2547163
Appl Environ Microbiol. 2008 Jun;74(11):3419-25
18408063
Bioessays. 1995 Aug;17(8):733-41
7661854
J Biol Chem. 2000 Nov 3;275(44):34609-18
10924523
J Mol Biol. 1982 Oct 15;161(1):57-76
6818353
Mol Cell. 2008 Feb 15;29(3):291-301
18280235
Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8376-80
11459978
EMBO J. 2001 Feb 15;20(4):924-32
11179236
Nucleic Acids Res. 2000 Mar 1;28(5):1067-77
10666445
Gene. 1995 Dec 1;166(1):177-8
8529886
Mol Microbiol. 2005 Sep;57(6):1664-75
16135232
Nat Rev Mol Cell Biol. 2006 May;7(5):335-46
16612326
Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11627-32
11573000
Annu Rev Biochem. 2005;74:283-315
15952889
EMBO J. 1998 Oct 1;17(19):5822-31
9755182
Bioessays. 2000 Nov;22(11):997-1006
11056476
Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):3936-41
10737770
J Biol Chem. 1995 Dec 8;270(49):29563-9
7493999
J Biol Chem. 2001 Dec 14;276(50):47185-94
11572866
Mol Cell. 2000 Sep;6(3):573-82
11030337
Mol Microbiol. 2006 Jan;59(2):460-74
16390442
Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
7984417