The telomeric GGGTTA repeats of Trypanosoma brucei contain the hypermodified base J in both strands

We have previously shown that nuclear DNA of bloodstream form Trypanosoma brucei contains a novel base β-glucosyl-hydroxymethyluracil, called J. Base J is enriched in minichromosome fractions but not in the minichromosome internal repeats, suggesting the association of J with telomeric DNA. To test whether J is present in the long telomeric (GGGTTA)(n) repeat arrays, which are 2-26 kb in T.brucei, we have purified these arrays both by hybrid selection and by isolating 2-26 kb fragments from DNA digested with multiple restriction enzymes. We find that in purified telomeric repeats ~ 13% of T is replaced by J, compared to 0.8% in total DNA, and we estimate that ~ 50% of the total J is in these repeats. Highly purified complementary strands of the repeats were obtained by alkaline CsCI equilibrium centrifugation, In the (TAACCC)(n) strand 14% of T was replaced by J. In the (GGGTTA)(n) strand ~ 36% of the second T was replaced by J; the first T was not detectably replaced. Modified bases have not been found in telomeric repeats before. How the bulky base J affects telomere function and structure in bloodstream form trypanosomes remains to be determined.