Expanded CTG triplet blocks from the myotonic dystrophy gene create the strongest known natural nucleosome positioning elements

Abstract

Expanded blocks of repeating nucleotide triplets have been found in or near genes associated with several human diseases. In the case of myotonic dystrophy, a block of repeating CTG trinucleotides is located downstream of the gene, and expansions of this block to repeats of n = 100 or more are frequently found in afflicted individuals. Using electron microscopy, we recently demonstrated that these blocks form unusually stable nucleosomes. Here, competitive nucleosome reconstitution was employed to measure the energetics of nucleosome formation over CTG repeat blocks of n = 75 and n = 130. These values were compared to the Xenopus borealis somatic 5S RNA gene, previously one of the strongest known natural nucleosome positioning elements. It is shown that DNA fragments containing 75 and 130 CTG repeats are 6 and 9 times stronger in nucleosome formation, respectively, than the 5S RNA gene. These findings suggest that expanded CTG blocks may profoundly alter local chromatin structure.