Anomalous rapid electrophoretic mobility of DNA containing triplet repeats associated with human disease genes.
Eight human genetic diseases have been associated with the expansion of CTG or CGG triplet repeats. The molecular etiology behind expansion is unknown but may involve participation of an unusual DNA structure in replication, repair, or recombination. We show that DNA fragments containing CTG triplet repeats derived from the human myotonic dystrophy gene migrate up to 20% faster than expected in nondenaturing polyacrylamide gels, suggesting the presence of an unusual DNA helix structure within the CTG triplet repeats. The anomalous migration is dependent upon the number of triplet repeats, the length of the flanking DNA, and the percentage and temperature of the polyacrylamide. The effect could be reduced by the addition of actinomycin D. Applying a reptation model for electrophoresis, the results are consistent with a 20% increase in persistence length of the DNA. PCR products containing CTG or CGG repeats from the spinocerebellar ataxia type I gene (SCA1) or the fragile X FMR1 gene, respectively, also showed higher electrophoretic mobility. These are the first sequences of defined length for which a dramatic increase in mobility can be attributed to sequence-dependent structural elements in DNA.
Chastain
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PD
Department of Biochemistry and Biophysics, Texas A&M University, Houston 77030-3303, USA.
Eichler
E E
EE
Kang
S
S
Nelson
D L
DL
Levene
S D
SD
Sinden
R R
RR
D000067696:Ataxin-1; D000067528:Ataxins; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003609:Dactinomycin / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D051860:Fragile X Messenger Ribonucleoprotein 1; D005600:Fragile X Syndrome / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009223:Myotonic Dystrophy / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D013132:Spinocerebellar Degenerations / Q000235:genetics*
D000067696:Ataxin-1; D000067528:Ataxins; D004247:DNA; D003609:Dactinomycin; D051860:Fragile X Messenger Ribonucleoprotein 1; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C000593749:ATXN1 protein, human; C497683:FMR1 protein, human