Abstract
gamma delta, a prokaryotic transposon, encodes a transposase that is essential for its transposition. We show here, by DNase I protection experiments, that purified gamma delta transposase binds at the transposon's inverted repeats (IRs). Immediately adjacent to each transposase binding site (and within gamma delta DNA) we have identified a binding site for an additional protein factor, the Escherichia coli-encoded integration host factor (IHF). The binding of transposase and IHF to these adjacent sites is mutually cooperative. An IHF binding-site was also found in the original target DNA, just outside one of the ends of gamma delta. The affinity of IHF for this flanking site is reduced by transposase. These results demonstrate that gamma delta transposase binds at the IRs of gamma delta, and suggest that IHF may be involved in forming a transposase-DNA complex and/or influencing the target site selection during the transposition of gamma delta.
MeSH Terms
Adenosine Triphosphate/metabolism
Bacterial Proteins/metabolism
Base Sequence
DNA Transposable Elements
DNA, Bacterial/metabolism
DNA-Binding Proteins/metabolism
Escherichia coli/genetics,metabolism
Integration Host Factors
Molecular Sequence Data
Nucleotidyltransferases/metabolism
Protein Binding
Transposases
Chemicals
Bacterial Proteins
DNA Transposable Elements
DNA, Bacterial
DNA-Binding Proteins
Integration Host Factors
Adenosine Triphosphate
Nucleotidyltransferases
Transposases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Wiater L A
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven 06510.
Grindley N D
References (27)
27 references, click to expand
-
Primary structure of the hip gene of Escherichia coli and of its product, the beta subunit of integration host factor.
J Mol Biol. 1985 May 25;183(2):117-28
PMID: 3159903
-
ATP-dependent specific binding of Tn3 transposase to Tn3 inverted repeats.
Nature. 1985 Apr 11-17;314(6011):556-8
PMID: 2986006
-
Transpositional recombination in prokaryotes.
Annu Rev Biochem. 1985;54:863-96
PMID: 2992361
-
DNA sequence at the end of IS1 required for transposition.
Nature. 1985 Oct 3-9;317(6036):458-60
PMID: 2995832
-
A defined system for the DNA strand-transfer reaction at the initiation of bacteriophage Mu transposition: protein and DNA substrate requirements.
Proc Natl Acad Sci U S A. 1985 Nov;82(22):7570-4
PMID: 2999771
-
Identification of the DNA sequence required for transposition immunity of the gamma delta sequence.
J Bacteriol. 1987 Sep;169(9):4388-90
PMID: 3040693
-
Mechanism of bacteriophage mu transposition.
Annu Rev Genet. 1986;20:385-429
PMID: 3028246
-
Host protein requirements for in vitro site-specific DNA inversion.
Cell. 1986 Aug 15;46(4):531-9
PMID: 3524854
-
Tn10 transposition and circle formation in vitro.
Cell. 1987 Oct 9;51(1):101-11
PMID: 2820584
-
Escherichia coli integration host factor binds specifically to the ends of the insertion sequence IS1 and to its major insertion hot-spot in pBR322.
J Mol Biol. 1987 May 20;195(2):261-72
PMID: 2821273
-
Histonelike proteins of bacteria.
Microbiol Rev. 1987 Sep;51(3):301-19
PMID: 3118156
-
Specific binding of transposase to terminal inverted repeats of transposable element Tn3.
Proc Natl Acad Sci U S A. 1987 Dec;84(23):8220-4
PMID: 2825182
-
Inhibition of TnA translocation by TnA.
J Bacteriol. 1977 Jan;129(1):407-14
PMID: 318647
-
A new method for sequencing DNA.
Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4
PMID: 265521
-
Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
J Bacteriol. 1978 Jun;134(3):1141-56
PMID: 149110
-
DNAse footprinting: a simple method for the detection of protein-DNA binding specificity.
Nucleic Acids Res. 1978 Sep;5(9):3157-70
PMID: 212715
-
Transposition of the Escherichia coli insertion element gamma generates a five-base-pair repeat.
Proc Natl Acad Sci U S A. 1979 Oct;76(10):4882-6
PMID: 388421
-
Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9.
Nature. 1979 Dec 20-27;282(5741):864-9
PMID: 390403
-
DNA sequence analysis of the transposon Tn3: three genes and three sites involved in transposition of Tn3.
Cell. 1979 Dec;18(4):1153-63
PMID: 391406
-
Purification and properties of the Escherichia coli protein factor required for lambda integrative recombination.
J Biol Chem. 1981 Sep 10;256(17):9246-53
PMID: 6267068
-
Transposon-specified site-specific recombination.
Proc Natl Acad Sci U S A. 1982 Jan;79(1):46-50
PMID: 6275390
-
Identification of a transposon Tn3 sequence required for transposition immunity.
Proc Natl Acad Sci U S A. 1983 Nov;80(22):6765-9
PMID: 6316324
-
Mechanism of transposition of bacteriophage Mu: polarity of the strand transfer reaction at the initiation of transposition.
Cell. 1984 Dec;39(2 Pt 1):395-404
PMID: 6094017
-
E. coli integration host factor binds to specific sites in DNA.
Cell. 1984 Dec;39(3 Pt 2):707-16
PMID: 6096022
-
Primary structure of the himA gene of Escherichia coli: homology with DNA-binding protein HU and association with the phenylalanyl-tRNA synthetase operon.
Cold Spring Harb Symp Quant Biol. 1984;49:691-8
PMID: 6397321
-
The phi 80 and P22 attachment sites. Primary structure and interaction with Escherichia coli integration host factor.
J Biol Chem. 1985 Apr 10;260(7):4468-77
PMID: 2984205
-
Sequence of the Escherichia coli pheST operon and identification of the himA gene.
J Bacteriol. 1985 Aug;163(2):787-91
PMID: 2991205