Effects of sequence on repeat expansion during DNA replication.
Small DNA repeat tracts are located throughout the human genome. The tracts are unstable, and expansions of certain repeat sequences cause neuromuscular disease. DNA expansions appear to be associated with lagging-strand DNA synthesis and DNA repair. At some sites of repeat expansion, e.g. the myotonic dystrophy type 2 (DM2) tetranucleotide repeat expansion site, more than one repeat tract with similar sequences lie side by side. Only one of the DM2 repeat tracts, however, is found to expand. Thus, DNA base sequence is a possible factor in repeat tract expansion. Here we determined the expansion potential, during DNA replication by human DNA polymerase beta, of several tetranucleotide repeat tracts in which the repeat units varied by one or more bases. The results show that subtle changes, such as switching T for C in a tetranucleotide repeat, can have dramatic consequences on the ability of the nascent-strand repeat tract to expand during DNA replication. We also determined the relative stabilities of self-annealed 100mer repeats by melting-curve analysis. The relative stabilities did not correlate with the relative potentials of the analogous repeats for expansion during DNA replication, suggesting that hairpin formation is not required for expansion during DNA replication.
Heidenfelder
Brooke L
BL
Lineberger Comprehensive Cancer Center, Department of Biochemistry and Biophysics, University of North Carolina Medical School, Chapel Hill, NC 27599-7295, USA.
Topal
Michael D
MD
D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D019951:DNA Polymerase beta / Q000378:metabolism; D042622:DNA Repeat Expansion / Q000235:genetics*; D004261:DNA Replication; D042822:Genomic Instability / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D009223:Myotonic Dystrophy / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D013816:Thermodynamics
D004247:DNA; D019951:DNA Polymerase beta