Anomalous rapid electrophoretic mobility of DNA containing triplet repeats associated with human disease genes

P. D. Chastain, E. E. Eichler, Seong Man Kang, D. L. Nelson, S. D. Levene, R. R. Sinden

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)16125-16131
Number of pages7
JournalBiochemistry
Volume34
Issue number49
DOIs
Publication statusPublished - 1995 Dec 29
Externally publishedYes

Fingerprint

Trinucleotide Repeats
Electrophoretic mobility
Genes
DNA
Spinocerebellar Ataxias
Recombinational DNA Repair
Myotonic Dystrophy
Inborn Genetic Diseases
Medical Genetics
Dactinomycin
Electrophoresis
Repair
Polymerase Chain Reaction
Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

Anomalous rapid electrophoretic mobility of DNA containing triplet repeats associated with human disease genes. / Chastain, P. D.; Eichler, E. E.; Kang, Seong Man; Nelson, D. L.; Levene, S. D.; Sinden, R. R.

In: Biochemistry, Vol. 34, No. 49, 29.12.1995, p. 16125-16131.

Research output: Contribution to journalArticle

Chastain, P. D. ; Eichler, E. E. ; Kang, Seong Man ; Nelson, D. L. ; Levene, S. D. ; Sinden, R. R. / Anomalous rapid electrophoretic mobility of DNA containing triplet repeats associated with human disease genes. In: Biochemistry. 1995 ; Vol. 34, No. 49. pp. 16125-16131.
@article{0a0645f9a6114ec08f42d364d2aadde7,
title = "Anomalous rapid electrophoretic mobility of DNA containing triplet repeats associated with human disease genes",
abstract = "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.",
author = "Chastain, {P. D.} and Eichler, {E. E.} and Kang, {Seong Man} and Nelson, {D. L.} and Levene, {S. D.} and Sinden, {R. R.}",
year = "1995",
month = "12",
day = "29",
doi = "10.1021/bi00049a027",
language = "English",
volume = "34",
pages = "16125--16131",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "49",

}

TY - JOUR

T1 - Anomalous rapid electrophoretic mobility of DNA containing triplet repeats associated with human disease genes

AU - Chastain, P. D.

AU - Eichler, E. E.

AU - Kang, Seong Man

AU - Nelson, D. L.

AU - Levene, S. D.

AU - Sinden, R. R.

PY - 1995/12/29

Y1 - 1995/12/29

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0028846524&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028846524&partnerID=8YFLogxK

U2 - 10.1021/bi00049a027

DO - 10.1021/bi00049a027

M3 - Article

C2 - 8519769

AN - SCOPUS:0028846524

VL - 34

SP - 16125

EP - 16131

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 49

ER -