PKD1 intron 21

Triplex DNA formation and effect on replication

Hiren P. Patel, Lu Lu, Richard T. Blaszak, John Bissler

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Although autosomal dominant polycystic kidney disease is transmitted in an autosomal dominant fashion, there is evidence that the pathophysiology of cystogenesis involves a second hit somatic mutation superimposed upon the inherited germline mutation within the renal tubule cells. The polypurine-polypyrimidine (Pu·Py) tract of PKD1 intron 21 may play a role in promoting somatic mutations. To better characterize this tract and to evaluate its potential to participate in mutagenesis, we investigated the thermodynamics of intramolecular triplex formation by 15 Pu·Py mirror repeat tracts from PKD1 intron 21 by 2D gel electrophoresis. We demonstrate that intramolecular triplexes form with modest superhelical tensions for all the tracts examined. Primer extension studies demonstrated significant polymerase arrest within the Pu·Py tracts in one direction of replication only. We found correlation between polymerization arrest and both the potential length of the triplex and superhelical tension of intramolecular triplex formation. The presence of a Pu·Py tract also led to a replication blockade and double-strand breakage using an SV40 in vitro replication assay with HeLa cell extracts. During DNA replication, the G-rich template of the PKD1 Pu·Py tracts may form a triplex structure with the nascent strand, thereby blocking replication and potentially leading to recombination and mutation.

Original languageEnglish (US)
Pages (from-to)1460-1468
Number of pages9
JournalNucleic acids research
Volume32
Issue number4
DOIs
StatePublished - Jul 14 2004

Fingerprint

Introns
Mutation
Autosomal Dominant Polycystic Kidney
Germ-Line Mutation
Electrophoresis, Gel, Two-Dimensional
Cell Extracts
DNA Replication
HeLa Cells
Thermodynamics
Mutagenesis
Polymerization
Genetic Recombination
Kidney
triplex DNA
Direction compound
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

PKD1 intron 21 : Triplex DNA formation and effect on replication. / Patel, Hiren P.; Lu, Lu; Blaszak, Richard T.; Bissler, John.

In: Nucleic acids research, Vol. 32, No. 4, 14.07.2004, p. 1460-1468.

Research output: Contribution to journalArticle

Patel, Hiren P. ; Lu, Lu ; Blaszak, Richard T. ; Bissler, John. / PKD1 intron 21 : Triplex DNA formation and effect on replication. In: Nucleic acids research. 2004 ; Vol. 32, No. 4. pp. 1460-1468.
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