| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 268, Issue 5, 3683-3689, Feb, 1993
G Pan, K Luetke, CD Juby, R Brousseau and P Sadowski
The FLP protein of the 2-microns plasmid of Saccharomyces cerevisiae is a
conservative site-specific recombinase that is involved in the
amplification of the plasmid. This recombination reaction proceeds via the
covalent attachment of the protein to the 3'-phosphoryl group at the site
of the breaks through a phosphotyrosine linkage. We have recently developed
an assay that measures FLP-mediated strand ligation independent of
FLP-mediated cleavage and covalent attachment to the DNA. The substrate for
ligation was produced by FLP-induced cleavage of the FLP recognition site
followed by digestion with Pronase and was shown to contain (at least) a
tyrosine residue at the 3'-PO4 terminus adjacent to the FLP cleavage sites.
We have now synthesized artificial substrates that bear a tyrosine residue
on the 3'-PO4 of an appropriate oligonucleotide and find that this
substrate is ligated as efficiently as the previous ligation substrates
that were isolated after FLP cleavage of the substrate. Analogous
substrates for other members of the integrase family of recombinases
(lambda integrase protein, P1-Cre protein) as well as for mammalian
topoisomerase I are also active as ligation substrates with their cognate
protein. This class of activated substrates should be useful in the study
of breakage and reunion reactions involving DNA.
Ligation of synthetic activated DNA substrates by site-specific recombinases and topoisomerase I
Department of Molecular and Medical Genetics, University of Toronto, Canada.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  C. D. Claeboe, R. Gao, and S. M. Hecht 3'-Modified oligonucleotides by reverse DNA synthesis Nucleic Acids Res., October 1, 2003; 31(19): 5685 - 5691. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. D. Bromberg, C. Hendricks, A. B. Burgin, and N. Osheroff Human Topoisomerase IIalpha Possesses an Intrinsic Nucleic Acid Specificity for DNA Ligation. USE OF 5' COVALENTLY ACTIVATED OLIGONUCLEOTIDE SUBSTRATES TO STUDY ENZYME MECHANISM J. Biol. Chem., August 16, 2002; 277(34): 31201 - 31206. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Woodfield, C. Cheng, S. Shuman, and A. B. Burgin Vaccinia topoisomerase and Cre recombinase catalyze direct ligation of activated DNA substrates containing a 3'-para-nitrophenyl phosphate ester Nucleic Acids Res., September 1, 2000; 28(17): 3323 - 3331. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. R. Knudsen, J. Lee, M. Lisby, O. Westergaard, and M. Jayaram Alcoholysis and Strand Joining by the Flp Site-specific Recombinase. MECHANISTICALLY EQUIVALENT REACTIONS MEDIATED BY DISTINCT CATALYTIC CONFIGURATIONS J. Biol. Chem., August 21, 1998; 273(34): 22028 - 22036. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-D. Zhu and P. D. Sadowski The Role of Single-stranded DNA in Flp-mediated Strand Exchange J. Biol. Chem., February 27, 1998; 273(9): 4921 - 4927. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. C. Shaikh and P. D. Sadowski The Cre Recombinase Cleaves the lox Site in trans J. Biol. Chem., February 28, 1997; 272(9): 5695 - 5702. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X.-D. Zhu and P. D. Sadowski Cleavage-dependent Ligation by the FLP Recombinase J. Biol. Chem., September 29, 1995; 270(39): 23044 - 23054. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
