| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print January 22, 2002
Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706
Corresponding Author: reznikoff{at}biochem.wisc.edu
This work identifies novel structure-function relationships between Tn5 transposase (Tnp) and its DNA recognition sequence. The Tn5 Tnp-DNA co-crystal structure revealed the protein-DNA contacts of the post-cleavage complex (1). One of the most striking features of this complex is the rotation of thymine 2 (T2) away from the DNA helix and into a pocket within Tnp. This interaction appears similar to the "base flipping" phenomenon found in many DNA repair enzymes such as T4 endonuclease V and uracil DNA glycosylase (2). To study the biochemical significance of this phenomenon, we mutated the Tnp residues proposed to be involved in stabilizing this interaction and removed the T2 nucleotide to examine which steps in the transposition reaction require T2-Tnp interactions. From this work we have determined that stacking interactions between T2 and Tnp are critical for efficient transposition in vitro. In addition, our results suggested that T2-Tnp interactions facilitate hairpin formation and hairpin resolution, primarily through base stacking, and that T2 plays a direct role in the strand transfer process.
J. Biol. Chem, 10.1074/jbc.M111119200
Submitted on November 20, 2001
Revised on January 9, 2002
Accepted on January 18, 2002
Mutational analysis of the base flipping event found in Tn5 transposition
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  S. Vaezeslami, R. Sterling, and W. S. Reznikoff Site-Directed Mutagenesis Studies of Tn5 Transposase Residues Involved in Synaptic Complex Formation J. Bacteriol., October 15, 2007; 189(20): 7436 - 7441. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Bischerour and R. Chalmers Base-flipping dynamics in a DNA hairpin processing reaction Nucleic Acids Res., April 4, 2007; (2007) gkm186v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. J. Grundy, J. E. Hesse, and M. Gellert Requirements for DNA hairpin formation by RAG1/2 PNAS, February 27, 2007; 104(9): 3078 - 3083. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-H. Saariaho and H. Savilahti Characteristics of MuA transposase-catalyzed processing of model transposon end DNA hairpin substrates Nucleic Acids Res., June 6, 2006; 34(10): 3139 - 3149. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Steiniger, C. D. Adams, J. F. Marko, and W. S. Reznikoff Defining characteristics of Tn5 Transposase non-specific DNA binding. Nucleic Acids Res., January 1, 2006; 34(9): 2820 - 2832. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. R. Arkhipova and M. Meselson Diverse DNA transposons in rotifers of the class Bdelloidea PNAS, August 16, 2005; 102(33): 11781 - 11786. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Kobryn, A. B. Burgin, and G. Chaconas Uncoupling the Chemical Steps of Telomere Resolution by ResT J. Biol. Chem., July 22, 2005; 280(29): 26788 - 26795. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. S. Reznikoff, S. R. Bordenstein, and J. Apodaca Comparative Sequence Analysis of IS50/Tn5 Transposase J. Bacteriol., December 15, 2004; 186(24): 8240 - 8247. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Bankhead and G. Chaconas Mixing active-site components: A recipe for the unique enzymatic activity of a telomere resolvase PNAS, September 21, 2004; 101(38): 13768 - 13773. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Ason and W. S. Reznikoff A high-throughput assay for Tn5 Tnp-induced DNA cleavage Nucleic Acids Res., June 16, 2004; 32(10): e83 - e83. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Peterson and W. Reznikoff Tn5 Transposase Active Site Mutations Suggest Position of Donor Backbone DNA in Synaptic Complex J. Biol. Chem., January 10, 2003; 278(3): 1904 - 1909. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
