Site-specific cleavage of single-stranded and double-stranded DNA sequences by oligodeoxyribonucleotides covalently linked to an intercalating agent and an EDTA-Fe chelate

Boidot-Forget, Marc; Chassignol, Marcel; Takasugi, Masashi; Thuong, Nguyen T.; Hélène, Claude

doi:10.1016/0378-1119(88)90163-1

Cited by 55 publications

(30 citation statements)

References 21 publications

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“…Addition of 1 mM spermine or increasing NaCl concentration to 1 M allowed cleavage to occur. These results are in qualitative agreement with those previously reported either with photocrosslinking agents (13,14) or with Fe-EDTA-oligonucleotide complexes (6,17,18).…”

supporting

confidence: 83%

“…Work from our laboratory has shown that photocrosslinking reactions could be induced on each strand of the double helix and that crosslinks could be converted into strand breaks upon alkaline treatment (13,14). The cleavage reaction induced by Fe-EDTA can be targeted to duplex DNA (6,17,18) as well as the crosslinking reaction mediated by a nitrogen mustard (19). The binding of homopyrimidine oligonucleotides to duplex DNA can be monitored by footprinting studies (20) or by gel retardation assays (21).…”

mentioning

confidence: 99%

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Sequence-specific recognition and cleavage of duplex DNA via triple-helix formation by oligonucleotides covalently linked to a phenanthroline-copper chelate.

François

Saison‐Behmoaras

Barbier

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

154

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Homopyrimidine oligodeoxynucleotides recognize the major groove of the DNA double helix at homopurine-homopyrimidine sequences by forming local triple helices. Phenanthroline was covalently attached to the 5' end of an 1i-mer homopyrimidine oligonucleotide of sequence d(TT-TCCTCCTCT). Simian virus 40 DNA, which contains a single target site for this oligonucleotide, was used as a substrate for the phenanthroline-oligonucleotide conjugate. In the presence of copper ions and a reducing agent, a single specific doublestrand cleavage site was observed at 20'C by agarose gel electrophoresis. The efficiency of double-strand cleavage was >70% at 20'C and pH 7.4. Secondary cleavage sites were observed when binding of the oligonucleotide to mismatched sequences was allowed to take place at low temperature. The exact location of the cleavage sites was determined by polyacrylamide gel electrophoresis of denatured fragments by using both simian virus 40 DNA and a synthetic DNA fragment containing the target sequence. The asymmetric distribution of the cleavage sites on the two strands revealed that the cleavage reaction took place in the minor groove even though the phenanthroline linker was located in the major groove. Linkers of different lengths were used to tether phenanthroline to the oligonucleotide and their relative efficacies of DNA cleavage were compared. Based on these comparative studies and on model building, it is proposed that the phenanthroline ring carried by the oligonucleotide intercalates from the major groove and that copper chelation locks the complex in place from within the minor groove where the cleavage reaction occurs.

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supporting

confidence: 83%

mentioning

confidence: 99%

Sequence-specific recognition and cleavage of duplex DNA via triple-helix formation by oligonucleotides covalently linked to a phenanthroline-copper chelate.

François

Saison‐Behmoaras

Barbier

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

154

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“…Benzalkonium chloride (B-1383, Sigma) is a cationic surface-active agent also used in vaginal spermicides. Proflavine (3,6-acridinamine, P-4646, Sigma), which is used as a topical antiseptic, has been shown to terminate DNA synthesis [49]. Chloroquine (generously provided by Dr. Yun-Yen Tsong, The Population Council, New York, NY) is a quinone analogue with antimalarial, antiamebic, and antirheumatic properties that is known for its suppressive effect on the immune system in lupus erythematosus.…”

Section: Reagentsmentioning

confidence: 99%

Studies of Adhesion of Lymphocytic Cells: Implications for Sexual Transmission of Human Immunodeficiency Virus1

Pearce-Pratt

Phillips

1993

Biology of Reproduction

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Considerable evidence suggests that sexual transmission of human immunodeficiency virus (HIV) is mediated via mononuclear cells that can infect epithelia of the genital tract. We describe here an in vitro model that can be used to examine the mechanism of cell-to-cell transmission of this virus. We have employed the system to identify agents that may be effective in a vaginal formulation to prevent HIV transmission via sexual contact. We have previously shown that chronically HIV-infected mononucleocytes can infect CD4-negative epithelial monolayers in the following manner: adhesion, via multiple microvilli, of HIV-infected mononucleocyte-derived cells to epithelial monolayers activates rapid virion secretion. Virions are then shed from the attached surface of the infected lymphocyte into a partially enclosed, microvilli-laden space between the cells. The shedding results in uptake of the virus and epithelial cell infection as demonstrated by ultrastructural examination and in vitro virological techniques. In this report, we present evidence from time-lapse films that HIV-infected lymphocytes adhere to the epithelium for a few minutes and then shift position to another site on the epithelium. As a result, one infected lymphocyte appears to be able to sequentially infected several cells of the epithelial monolayer. Using a fluorescence-based cell-cell adhesion assay to examine the effect of seminal fluid and a variety of chemical compounds on lymphocyte-to-epithelial adherence, we found that seminal fluid significantly increases the number of lymphocytes adhering to epithelia. This suggests that semen can serve as an effective medium for cell-cell transmission of HIV. On the other hand, sulfated polysaccharides and glutathione effectively inhibit cell-cell adhesion. Since the cell-cell adhesion step is critical to epithelial cell infection by HIV, these results suggest that anti-cell adhesion compounds may be effective in a vaginal formulation to reduce the probability of HIV infection.

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“…27 for review). It is also possible to induce strand breaks in the target nucleic acid by covalent linkage of metal complexes that generate hydroxyl radicals (27)(28)(29)(30)(31)(32)(33). Here we describe a photoactive system that can be used to photocrosslink the oligonucleotide to its complementary sequence.…”

Section: Discussionmentioning

confidence: 99%

Sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of DNA via triple-helix formation.

Praseuth

Perrouault

Doan

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

217

119

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A photocrosslinking reagent (p-azidophenacyl) was covalently linked to an octathymidylate synthesized with either the natural ((3) anomer of thymidine or the synthetic (a) anomer. The oligothymidylate was further substituted by an acridine derivative to stabilize the hybrid formed with a complementary octadeoxyadenylate sequence via intercalation. A single-stranded 27-mer containing a (dA)8 sequence and a 27-mer duplex containing a (dA-dT)8 sequence were used as targets. Upon UV irradiation, photocrosslinking of the octathymidylate to its target sequence was observed, generating bands that migrated more slowly in denaturing gels. In the 27-mer duplex, both strands were photocrosslinked to the octathymidylate. Upon alkaline treatment of the irradiated samples, cleavage of the 27-mers was observed at specific sites. These reactions were analyzed at different salt concentrations. The location of the cleavage sites allowed us to demonstrate the following. (i) Both a and (3 oligothymidylates can recognize a DNA double helix containing an oligo(dA)-oligo(dT) sequence; the oligothymidylate binds to the major groove of DNA in a parallel orientation with respect to the adenine-containing strand of the DNA double helix. (ii) a oligothymidylates form helices with a complementary singlestranded oligodeoxyadenylate; the two strands have a parallel orientation independently of whether or not an intercalating agent is attached to the oligothymidylate. (iii) At low salt concentration, (3 oligothymidylates form a double helix with an oligodeoxyadenylate in which, as expected, the two strands are antiparallel; at high salt concentration, a triple helix is formed in which the second oligothymidylate is oriented parallel to the adenine-containing strand. These results show that it is possible to recognize an oligopurine-oligopyrimidine sequence in a DNA double helix via local triple-helix formation and to target photochemical reactions to specific sequences in both double-stranded and single-stranded nucleic acids.

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Site-specific cleavage of single-stranded and double-stranded DNA sequences by oligodeoxyribonucleotides covalently linked to an intercalating agent and an EDTA-Fe chelate

Cited by 55 publications

References 21 publications

Sequence-specific recognition and cleavage of duplex DNA via triple-helix formation by oligonucleotides covalently linked to a phenanthroline-copper chelate.

Sequence-specific recognition and cleavage of duplex DNA via triple-helix formation by oligonucleotides covalently linked to a phenanthroline-copper chelate.

Studies of Adhesion of Lymphocytic Cells: Implications for Sexual Transmission of Human Immunodeficiency Virus1

Sequence-specific binding and photocrosslinking of alpha and beta oligodeoxynucleotides to the major groove of DNA via triple-helix formation.

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