Nucleic Acids Research, Vol 24, Issue 13 2476-2482, Copyright © 1996 by Oxford University Press
F van Leeuwen, ER Wijsman, E Kuyl-Yeheskiely, GA van der Marel, JH van Boom and P Borst
We have previously shown that nuclear DNA of bloodstream from Trypanosoma
brucei contains a novel base beta-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 approximately 13% of T is replaced by J, compared to 0.8%
in total DNA, and we estimate that approximately 50% of the total J is in
these repeats. Highly purified complementary strands of the repeats were
obtained by alkaline CsCl equilibrium centrifugation. In the (TAACCC)n
strand 14% of T was replaced by J. In the (GGGTTA)n strand approximately
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.
ARTICLES
The telomeric GGGTTA repeats of Trypanosoma brucei contain the hypermodified base J in both strands
Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
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