Antiserum to Z‐DNA

Malfoy, Bernard; Leng, Marc

doi:10.1016/0014-5793(81)80424-3

Cited by 61 publications

(25 citation statements)

References 26 publications

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“…Perturbing effects of anti-Z-DNA antibodies on the salt-induced B-Z transition of poly(dG-dC).poly(dG-dC) have been observed (Malfoy and Leng, 1981;Jovin et al, 1983). Revet et al (1984) observed that if anti-Z-DNA antibodies were bound to supercoiled plasmids in very low ionic strength, and the plasmids were then cut with a restriction enzyme, some antibodies could be visualized on the linear plasmids in the electron microscope, suggesting that Z-DNA had been stabilized in the plasmid through its interaction with the anti-Z-DNA antibody.…”

Section: Introductionmentioning

confidence: 92%

Anti-Z-DNA antibody binding can stabilize Z-DNA in relaxed and linear plasmids under physiological conditions.

Lafer¹,

Sousa²,

Rich³

1985

The EMBO Journal

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It is shown that anti-Z-DNA antibody binding can stabilize sequences of d(CG/GC)Q and d(CA/GT). in the Z-DNA conformation in a plasmid in the complete absence of supercoiling. This effect is quantitated by using antibody preparations of different affinities and varying concentrations. The d(CG/GC)n sequence can be stabilized under physiological conditions. This is the first demonstration that a region of Z-DNA can be stabilized by protein binding in a completely relaxed plasmid under physiological conditions. The antibody-Z-DNA complex in the relaxed plasmid is shown to be an equilibrium state and not a long-lived kinetic intermediate since specific binding of the antibody to linearized plasmids containing Z-forming sequences is observed.

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Section: Introductionmentioning

confidence: 92%

Anti-Z-DNA antibody binding can stabilize Z-DNA in relaxed and linear plasmids under physiological conditions.

Lafer¹,

Sousa²,

Rich³

1985

The EMBO Journal

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“…The discovery that Z-DNA is a strong immunogen (17,18) has paved the way to look for the occurrence of such DNA structures in more complicated systems by taking advantage of the high specificity of immunochemical methods. For example, polytene chromosomes of Drosophila bind antibodies specific for Z-DNA (19), suggesting that such left-handed DNA conformations occur as structural elements in vivo.…”

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confidence: 99%

Isolation of Z-DNA-containing plasmids.

Thomae¹,

Beck²,

Pohl³

1983

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

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A purified, monoclonal antibody, specific for the left-handed Z-form of poly(dG-dC), was coupled covalently to Sephacryl S-1000 beads. Such an antibody column provides a convenient method to isolate and purify those plasmid DNAs that contain Z-DNA from a large excess of other DNAs, RNA, etc. From a library of Escherichia coli DNA, cloned into the vector plasmid pUC-8, several recombinant plasmids were isolated, which bind to this antibody. Thus, E. coli contains sequences, which in "natural" negatively supercoiled DNA, adopt a left-handed Z-DNAlike conformation.Physicochemical studies of poly-and oligo (dC-dG (17), were used for cell fusion (22, 23). Cell supernatants were screened for precipitating 32P-labeled poly(dGdC) in 4 M NaCl. Seven monoclonal cell lines producing antibodies to Z-DNA were established (unpublished data). Antibodies (K/y2b) from ascites fluids, by using cell line Z-DI1, were purified by protein A-Sepharose (Pharmacia) chromatography (24, 25). Antibody concentration was determined, by using A280 = 1.4 for a solution at 1 mg/ml. l"I-Labeled antibodies with -10,000 cpm/ng were prepared by the Chloramin-T method (26).Antibody Column. Sephacryl S-1000 (superfine; Pharmacia) was activated with cyanogen bromide in sodium carbonate buffer (27) The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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“…The antibodies were elicited in rabbits immunized with poly(dGdC)poly(dG-dC) chemically modified with chlorodiethylenetriamineplatinum(II) chloride. The antibodies were found to bind to a number of synthetic polynucleotides in Z form and to form V DNA, but no crossreaction was detected with native or denatured DNA (calf thymus or Micrococcus luteus DNA), singlestranded or double-stranded RNA, deoxyguanosine, or deoxycytidine (14,29,30). The chromosome staining is abolished when the antibodies are first incubated with poly(dG-br5dC)poly(dGbr5dC) polymer, which has the Z conformation irrespective of the salt concentration (30).…”

mentioning

confidence: 99%

“…Antisera against Z-DNA were. elicited in rabbits immunized with poly(dGdC)-poly (dG-dC) having 12% of the bases modified with chlorodiethylenetriamineplatinum(II) chloride (29,30). It has been shown that this modified polynucleotide exists in the Z form under physiological conditions (31).…”

mentioning

confidence: 99%

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Z-DNA immunoreactivity in fixed metaphase chromosomes of primates.

Viegas‐Péquignot¹,

Derbin²,

Malfoy³

et al. 1983

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

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Antibodies against Z-DNA bind to fixed metaphase chromosomes of man and Cebus albifrons (Platyrrhini, Primate). By indirect immunofluorescence and indirect immunoperoxidase techniques, a heavy staining is detected in some segments of chromosomes of C. albifrons. These segments correspond to Rband-positive heterochromatin, which has a high G+C-base content. Euchromatin of human and Cebus chromosomes show a weak and heterogeneous staining that consistently reproduces an R-and T-banding pattern in both species. Because chromosome homologies previously were demonstrated between these distantly related species by chromosome banding, our results suggest that Z-DNA has been conserved during the course of primate evolution.Repeating sequences of alternating dC and dG deoxynucleotides can form the left-handed duplex DNA conformation, named Z-DNA, as demonstrated by x-ray diffraction studies of crystals of (dC-dG)n (1-3) and fibers of poly (dG-dC)-poly (dG-dC) ( (23, 24), the comparison of the staining patterns produced by antibodies to Z-DNA in euchromatin may be considered a check of the stability of Z-DNA conformation during chromosome evolution.In this work we show (i) that anti-Z-DNA antibodies bind to metaphase chromosomes of man and Cebus in a distinct and specific way and (ii) that an undisputable correspondence exists between the antibody binding and the chromosome banding patterns. In euchromatin, a weak but distinct and consistent staining is observed in the segments corresponding to the Rand T-bands, which are thought to be richer in G-C base pairs both in man and Cebus. Slight differences of intensity are observed from band to band: they are the same for all of the chromosomal segments of man and Cebus, which were found homologous by banding pattern and by gene mapping comparison (23)(24)(25). This fact underlines the stability of Z-DNA conformation during the evolution of mammalian chromosomes. Furthermore, the additional heterochromatic segments existing in Cebus chromosomes exhibit a consistent and very strong binding to anti-Z-DNA antibodies.MATERIAL AND METHODS Obtention of Metaphase Chromosomes. Metaphase cells were obtained from tissue cultures after skin biopsy of a female of C. albifrons held in captivity at the Museum National d'Histoire Naturelle, Paris.Metaphase cells from human lymphocytes were cultured according to usual methods (26). Cell cultures were treated by colchicine for 1 or 2 hr and then for 10 min with a hypotonic solution of diluted human serum (1:6, vol/vol). Cells were fixed in most of the experiments for at least 40 min with two different solutions (solution I, ethanol/chloroform/acetic acid, 6:3:1, vol/ vol; solution II, ethanol/acetic acid, 3:1, vol/vol). A prolonged fixation (45 min to 48 hr), with only the last fixative solution was used for some human cultures.The following banding methods were used to characterize the chromosomes of the specimen of C. albifrons: R-banding 5890 The publication costs of this article were defrayed in part by page charge payment. T...

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Antiserum to Z‐DNA

Cited by 61 publications

References 26 publications

Anti-Z-DNA antibody binding can stabilize Z-DNA in relaxed and linear plasmids under physiological conditions.

Anti-Z-DNA antibody binding can stabilize Z-DNA in relaxed and linear plasmids under physiological conditions.

Isolation of Z-DNA-containing plasmids.

Z-DNA immunoreactivity in fixed metaphase chromosomes of primates.

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