5-Hydroxymethyluracil in the DNA of a Dinoflagellate

During the characterization of DNA from the dinoflagellate Gyrodinium cohnii, a large discrepancy was detected between the estimation of guanine + cytosine content from the buoyant density of the DNA in CsCl (56.1% G+C) and from the midpoint (Tm) of its hyperchromicity induced by a thermal gradient (35.6% G+C). Composition analyses of32P-labeled nucleotides revealed an actual G+C content of 41.3%, and the presence of an unusual nucleotide amounting to about 37% of the expected thymidylate in unfractionated DNA-a feature that can explain the aberrant behavior of the DNA. The chromatographic properties of the unusual base and UV spectral analyses of the base and its corresponding nucleotide are consistent with its identification as hydroxymethyluracil. This base is not uniformly interspersed with thymine in the DNA. About 10% of Gyrodinium DNA is contributed by a fraction with low hydroxymethyluracil content, which behaves anomalously in Ag+-Cs2SO4density gradients but not in CsCl.