Human MSH2 binds to trinucleotide repeat DNA structures associated with neurodegenerative diseases.
The expansion of trinucleotide repeat sequences is associated with several neurodegenerative diseases. The mechanism of this expansion is unknown but may involve slipped-strand structures where adjacent rather than perfect complementary sequences of a trinucleotide repeat become paired. Here, we have studied the interaction of the human mismatch repair protein MSH2 with slipped-strand structures formed from a triplet repeat sequence in order to address the possible role of MSH2 in trinucleotide expansion. Genomic clones of the myotonic dystrophy locus containing disease-relevant lengths of (CTG)n x (CAG)n triplet repeats were examined. We have constructed two types of slipped-strand structures by annealing complementary strands of DNA containing: (i) equal numbers of trinucleotide repeats (homoduplex slipped structures or S-DNA) or (ii) different numbers of repeats (heteroduplex slipped intermediates or SI-DNA). SI-DNAs having an excess of either CTG or CAG repeats were structurally distinct and could be separated electrophoretically and studied individually. Using a band-shift assay, the MSH2 was shown to bind to both S-DNA and SI-DNA in a structure-specific manner. The affinity of MSH2 increased with the length of the repeat sequence. Furthermore, MSH2 bound preferentially to looped-out CAG repeat sequences, implicating a strand asymmetry in MSH2 recognition. Our results are consistent with the idea that MSH2 may participate in trinucleotide repeat expansion via its role in repair and/or recombination.
Pearson
C E
CE
Center for Genome Research, Institute of Biosciences and Technology in the Texas Medical Center, Texas A&M University, Houston 77030, USA. cpearson@ibt.tamu.edu
Ewel
A
A
Acharya
S
S
Fishel
R A
RA
Sinden
R R
RR
D001667:Binding, Competitive; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004260:DNA Repair; D004268:DNA-Binding Proteins; D006801:Humans; D051718:MutS Homolog 2 Protein; D009410:Nerve Degeneration / Q000235:genetics*; D009692:Nucleic Acid Heteroduplexes; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism*; D013379:Substrate Specificity; D018911:Trinucleotide Repeats
D004268:DNA-Binding Proteins; D009692:Nucleic Acid Heteroduplexes; D011518:Proto-Oncogene Proteins; D004247:DNA; C497172:MSH2 protein, human; D051718:MutS Homolog 2 Protein