Pausing of DNA synthesis in vitro at specific loci in CTG and CGG triplet repeats from human hereditary disease genes

Seong Man Kang, K. Ohshima, M. Shimizu, S. Amirhaeri, R. D. Wells

Research output: Contribution to journalArticle

160 Citations (Scopus)

Abstract

Several human hereditary neuromuscular disease genes are associated with the expansion of CTG or CGG triplet repeats. The DNA syntheses of CTG triplets ranging from 17 to 180 and CGG repeats from 9 to 160 repeats in length were studied in vitro. Primer extensions using the Klenow fragment of DNA polymerase I, the modified T7 DNA polymerase (Sequenase), or the human DNA polymerase β paused strongly at specific loci in the CTG repeats. The pausings were abolished by heating at 70 °C. As the length of the triplet repeats in duplex DNA, but not in single-stranded DNA, was increased, the magnitude of pausings increased. The location of the pause sites was determined by the distance between the site of primer hybridization and the beginning of the triplet repeats. CGG triplet repeats also showed similar, but not identical, patterns of pausings. These results indicate that appropriate lengths of the triplets adopt a non-B conformation(s) that blocks DNA polymerase progression; the resultant idling polymerase may catalyze slippages to give expanded sequences and hence provide the molecular basis for this non-Mendelian genetic process. These mechanisms, if present in human cells, may be related to the etiology of certain neuromuscular diseases such as myotonic dystrophy and Fragile X syndrome.

Original languageEnglish
Pages (from-to)27014-27021
Number of pages8
JournalJournal of Biological Chemistry
Volume270
Issue number45
DOIs
Publication statusPublished - 1995 Jan 1
Externally publishedYes

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Trinucleotide Repeats
Inborn Genetic Diseases
DNA-Directed DNA Polymerase
Genes
DNA Polymerase I
Neuromuscular Diseases
DNA
Genetic Phenomena
Fragile X Syndrome
Myotonic Dystrophy
Single-Stranded DNA
Heating
Conformations
Cells
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pausing of DNA synthesis in vitro at specific loci in CTG and CGG triplet repeats from human hereditary disease genes. / Kang, Seong Man; Ohshima, K.; Shimizu, M.; Amirhaeri, S.; Wells, R. D.

In: Journal of Biological Chemistry, Vol. 270, No. 45, 01.01.1995, p. 27014-27021.

Research output: Contribution to journalArticle

Kang, Seong Man ; Ohshima, K. ; Shimizu, M. ; Amirhaeri, S. ; Wells, R. D. / Pausing of DNA synthesis in vitro at specific loci in CTG and CGG triplet repeats from human hereditary disease genes. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 45. pp. 27014-27021.
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