Evidence of cis-acting factors in replication-mediated trinucleotide repeat instability in primate cells.
The mechanism of disease-associated trinucleotide repeat instability involves cis-acting factors (cis-elements) in the vicinity of the repeat, but the nature of these elements is unknown. One cis-element may be the location of the replication origin relative to the repeat. We have used an SV40 DNA replication system to investigate the effect of the location of replication initiation on (CTG)(n)*(CAG)(n) stability in primate cells. Depending on the distance between the SV40 replication origin and the repeat tract, templates with 79 repeats yield predominantly expansions or predominantly deletions or remain intact. All templates with 17 repeats are stable. Thus, cis-elements that affect the sites of Okazaki fragment initiation relative to the repeat are crucial determinants of instability. This model system recapitulates the bias for expansions observed in many of the diseases associated with trinucleotide repeats. Our results might explain the variable amounts of CTG/CAG instability that are observed in different chromosomal contexts.
Cleary
John D
JD
Program of Genetics & Genomic Biology, The Hospital for Sick Children, 555 University Avenue, Elm Wing 11-135, Toronto, Ontario M5G 1X8, Canada.
Nichol
Kerrie
K
Wang
Yuh-Hwa
YH
Pearson
Christopher E
CE
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D002875:Chromosomes / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D004261:DNA Replication / Q000235:genetics; D018598:Minisatellite Repeats; D008957:Models, Genetic; D016133:Polymerase Chain Reaction; D018741:Replication Origin; D013539:Simian virus 40 / Q000235:genetics; D018911:Trinucleotide Repeats