| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 276, Issue 21, 18018-18023, May 25, 2001
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Departments of Therapeutic Radiology and Genetics, Yale
University School of Medicine, New Haven, Connecticut
06520-8040
Triple helix-forming oligonucleotides (TFOs) can
bind to polypurine/polypyrimidine regions in DNA in a sequence-specific
manner. Triple helix formation has been shown to stimulate
recombination in mammalian cells in both episomal and chromosomal
targets containing direct repeat sequences. Bifunctional
oligonucleotides consisting of a recombination donor domain tethered to
a TFO domain were found to mediate site-specific recombination in an
intracellular SV40 vector target. To elucidate the mechanism of
triplex-induced recombination, we have examined the ability of
intermolecular triplexes to provoke recombination within plasmid
substrates in human cell-free extracts. An assay for reversion of a
point mutation in the supFG1 gene in the plasmid
pSupFG1/G144C was established in which recombination in the extracts
was detected upon transformation into indicator bacteria. A
bifunctional oligonucleotide containing a 30-nucleotide TFO domain
linked to a 40-nucleotide donor domain was found to mediate gene
correction in vitro at a frequency of 46 × 10
Triplex-induced Recombination in Human Cell-free
Extracts
DEPENDENCE ON XPA AND HsRad51*
5, at least 20-fold above background and
over 4-fold greater than the donor segment alone. Physical linkage of
the TFO to the donor was unnecessary, as co-mixture of separate TFO and
donor segments also yielded elevated gene correction frequencies. When
the recombination and repair proteins HsRad51 and XPA were depleted
from the extracts using specific antibodies, the triplex-induced
recombination was diminished, but was either partially or completely
restored upon supplementation with the purified HsRad51 or XPA
proteins, respectively. These results establish that triplex-induced,
intermolecular recombination between plasmid targets and short
fragments of homologous DNA can be detected in human cell extracts and
that this process is dependent on both XPA and HsRad51.
*
This work was supported by National Institutes of Health
Grants RO1GM54731 and RO1CA64186 (to P. M. G.) and R37GM33504
(to C. M. R.) and by a scholar award (to P. M. G.)
from the Leukemia and Lymphoma Society.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Depts. of Therapeutic
Radiology and Genetics, Yale University School of Medicine, P.O.
Box 208040, New Haven, CT 06520-8040. Tel.: 203-737-2788; Fax:
203-737-2630; E-mail: peter.glazer@yale.edu.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Duca, P. Vekhoff, K. Oussedik, L. Halby, and P. B. Arimondo The triple helix: 50 years later, the outcome Nucleic Acids Res., September 1, 2008; 36(16): 5123 - 5138. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Liu, R. S. Nairn, and K. M. Vasquez Processing of triplex-directed psoralen DNA interstrand crosslinks by recombination mechanisms Nucleic Acids Res., August 1, 2008; 36(14): 4680 - 4688. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. M. Kalish, M. M. Seidman, D. L. Weeks, and P. M. Glazer Triplex-induced recombination and repair in the pyrimidine motif Nucleic Acids Res., June 16, 2005; 33(11): 3492 - 3502. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. A. Rogers, M. Manoharan, P. Rabinovitch, D. C. Ward, and P. M. Glazer Peptide conjugates for chromosomal gene targeting by triplex-forming oligonucleotides Nucleic Acids Res., December 15, 2004; 32(22): 6595 - 6604. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. S. Bindra, P. J. Schaffer, A. Meng, J. Woo, K. Maseide, M. E. Roth, P. Lizardi, D. W. Hedley, R. G. Bristow, and P. M. Glazer Down-Regulation of Rad51 and Decreased Homologous Recombination in Hypoxic Cancer Cells Mol. Cell. Biol., October 1, 2004; 24(19): 8504 - 8518. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. G. Uil, H. J. Haisma, and M. G. Rots Therapeutic modulation of endogenous gene function by agents with designed DNA-sequence specificities Nucleic Acids Res., November 1, 2003; 31(21): 6064 - 6078. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. A. Rogers, K. M. Vasquez, M. Egholm, and P. M. Glazer Site-directed recombination via bifunctional PNA-DNA conjugates PNAS, December 24, 2002; 99(26): 16695 - 16700. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. B. Panigrahi, J. D. Cleary, and C. E. Pearson In Vitro (CTG){middle dot}(CAG) Expansions and Deletions by Human Cell Extracts J. Biol. Chem., April 12, 2002; 277(16): 13926 - 13934. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. D. Richardson, L. B. Augustin, B. T. Kren, and C. J. Steer Gene Repair and Transposon-Mediated Gene Therapy Stem Cells, March 1, 2002; 20(2): 105 - 118. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. P. Knauert and P. M. Glazer Triplex forming oligonucleotides: sequence-specific tools for gene targeting Hum. Mol. Genet., October 1, 2001; 10(20): 2243 - 2251. [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 |
