Sticky DNA, a long GAA.GAA.TTC triplex that is formed intramolecularly, in the sequence of intron 1 of the frataxin gene.
Friedreich's ataxia is caused by the massive expansion of GAA.TTC repeats in intron 1 of the frataxin (X25) gene. Our prior investigations showed that long GAA.TTC repeats formed very stable triplex structures which caused two repeat tracts to adhere to each other (sticky DNA). This process was dependent on negative supercoiling and the presence of divalent metal ions. Herein, we have investigated the formation of sticky DNA from plasmid monomers and dimers; sticky DNA is formed only when two tracts of sufficiently long (GAA.TTC)(n) (n = 59-270) are present in a single plasmid DNA and are in the direct repeat orientation. If the inserts are in the indirect (inverted) repeat orientation, no sticky DNA was observed. Furthermore, kinetic studies support the intramolecular nature of sticky DNA formation. Electron microscopy investigations also provide strong data for sticky DNA as a single long triplex. Hence, these results give new insights into our understanding of the capacity of sticky DNA to inhibit transcription and thereby reduce the level of frataxin protein as related to the etiology of Friedreich's ataxia.
Vetcher
Alexandre A
AA
Center for Genome Research, Institute of Biosciences and Technology, Texas A&M University, Texas Medical Center, Houston, Texas 77030-3303, USA.
Napierala
Marek
M
Iyer
Ravi R
RR
Chastain
Paul D
PD
Griffith
Jack D
JD
Wells
Robert D
RD
D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure; D019281:Dimerization; D007438:Introns; D033862:Iron-Binding Proteins / Q000235:genetics*; D007700:Kinetics; D008854:Microscopy, Electron; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000378:metabolism; D020033:Protein Isoforms; D000096923:Frataxin
D004247:DNA; D033862:Iron-Binding Proteins; D020033:Protein Isoforms; D000096923:Frataxin; C051802:triplex DNA