The purpose of this paper is to show that the antibodies to nucleic acids, to nucleosides or to DNA damaged by a physical or a chemical agent, are useful tools in the study of DNA damage and repair. The results obtained with antibodies to nucleosides, antibodies to nucleosides and DNA modified by chemical carcinogens emphasize the potential of immunological methods in three main areas, a) the sensitive detection and quantitation of adducts; b) the visualization of adducts in tissues, individual cells, and along the DNA double helix; c) the study of conformational changes of DNA induced by adducts.

Animals↗

PubMedDate not supplied

Z-DNA immunoreactivity in rat tissues.

Recently, it has been shown that natural and synthetic deoxynucleotide polymers can adopt a left-handed helical structure (termed Z-DNA) in appropriate conditions (see, for example, refs 1 and 2 and the references therein). In contrast to the more familiar right-handed B-DNA, Z-DNA is strongly immunogenic, and polyclonal and monoclonal antibodies against Z-DNA have been elicited. By using such antibodies, immunoreactivity for Z-DNA has been detected in the polytene chromosomes of two dipteran species, in the macronucleus of a ciliated protozoon, and in certain plant nuclei (cited in ref. 11). In view of the possible importance of Z-DNA as a genomic regulatory signal, it would be highly desirable to know whether Z-DNA also occurs in mammals. We have therefore initiated an immunohistochemical study of various rat tissues by using three antisera specific for Z-DNA, and the peroxidase-antiperoxidase technique for visualization of tissue-bound antibodies. Here we demonstrate that the nuclei of many, but not all, types of rat cells exhibit Z-DNA immunoreactivity, suggesting that Z-DNA may exist naturally in mammalian chromatin.

Animals↗

PubMedDate not supplied

Ring-opened alkylated guanine is not repaired in Z-DNA.

Since the discovery of Z-DNA by X-ray analysis of the alternated hexanucleotide d(C-G)3 crystals, numerous studies have shown that fragments of natural DNAs can adopt the Z conformation, topological constraints being a major factor stabilizing this conformation. Immunochemical assays using antibodies to Z-DNA provide strong evidence for the presence of Z fragments in chromosomes. The biological role of Z-DNA is not yet known, but it might be involved in gene regulation. Proteins which bind specifically to Z-DNA have been isolated and interactions between Z-DNA and several cellular proteins have been studied. The ability of DNA repair enzymes to maintain the genome's integrity is of major importance to the cell. On alkylation of DNA by chemical carcinogens such as dimethyl sulphate, methyl methanesulphonate, methylnitrosourea or methylnitrosoguanidine, the main target is the N7 of the guanosine residue, yielding 7-methylguanine (mG). In alkaline conditions, the imidazole ring of mG opens up, yielding the ring-opened form 2,6-diamino-4-oxo-5-methylformamidopyrimidine (rom7G); this lesion is a block to DNA replication. It occurs in vivo and is enzymatically removed by the DNA glycosylase. Here we report that the lesion is not excised when present in DNA in the left-handed Z conformation.

DNA Repair↗