Genomic DNA of Nostoc commune ( Cyanobacteria) became covalently modified during decades of desiccation. Amplification of gene loci from desiccated cells required pretreatment of DNA with N-phenacylthiazolium bromide, a reagent that cleaves DNA- and protein-linked advanced glycosylation end-products. DNA from 13 year desiccated cells did not show any higher levels of the commonly studied oxidatively modified DNA damage biomarkers 8-hydroxyguanine, 8-hydroxyadenine and 5-hydroxyuracil, compared to commercially available calf thymus DNA. Different patterns of amplification products were obtained with DNA from desiccated/ rehydrating cells and a liquid culture derived from the dried material, using the same set of primers. In contrast, a reproducible fingerprint was obtained, irrespective of time of rehydration of the DNA, using a primer (5'-GWCWATCGCC-3') based upon a highly iterated palindromic repeat sequence present in the genome. In vitro, the desiccation of cccDNA led to loss of supercoiling, aggregation, loss of resolution during agarose gel electrophoresis and loss of transformation and transfection efficiency. These changes were minimized when DNA was desiccated and stored in the presence of trehalose, a non-reducing disaccharide present in Nostoc colonies. The response of the N. commune genome to desiccation is different from the response of the genomes of cyanobacteria and Deinococcus radiodurans to ionizing radiation.
C1 Virginia Tech, Ctr Genom VIGEN, Blacksburg, VA 24061 USA.
Virginia Tech, Dept Chem, Blacksburg, VA 24061 USA.
Natl Inst Stand & Technol, DNA Technol Grp, Chem Sci & Technol Lab, Gaithersburg, MD 20899 USA.
Univ Maryland Baltimore Cty, Baltimore, MD 21250 USA.
Inonu Univ, Sch Med, Dept Pharmacol, Malatya, Turkey.