Non-B DNA conformations formed by long repeating tracts of myotonic dystrophy type 1, myotonic dystrophy type 2, and Friedreich's ataxia genes, not the sequences per se, promote mutagenesis in flanking regions.
The expansions of long repeating tracts of CTG.CAG, CCTG.CAGG, and GAA.TTC are integral to the etiology of myotonic dystrophy type 1 (DM1), myotonic dystrophy type 2 (DM2), and Friedreich's ataxia (FRDA). Essentially all studies on the molecular mechanisms of this expansion process invoke an important role for non-B DNA conformations which may be adopted by these repeat sequences. We have directly evaluated the role(s) of the repeating sequences per se, or of the non-B DNA conformations formed by these sequences, in the mutagenic process. Studies in Escherichia coli and three types of mammalian (COS-7, CV-1, and HEK-293) fibroblast-like cells revealed that conditions which promoted the formation of the non-B DNA structures enhanced the genetic instabilities, both within the repeat sequences and in the flanking sequences of up to approximately 4 kbp. The three strategies utilized included: the in vivo modulation of global negative supercoil density using topA and gyrB mutant E. coli strains; the in vivo cleavage of hairpin loops, which are an obligate consequence of slipped-strand structures, cruciforms, and intramolecular triplexes, by inactivation of the SbcC protein; and by genetic instability studies with plasmids containing long repeating sequence inserts that do, and do not, adopt non-B DNA structures in vitro. Hence, non-B DNA conformations are critical for these mutagenesis mechanisms.
Wojciechowska
Marzena
M
Institute of Biosciences and Technology, Center for Genome Research, Texas A&M University System Health Science Center, Houston, Texas 77030, USA.
Napierala
Marek
M
Larson
Jacquelynn E
JE
Wells
Robert D
RD
D000818:Animals; D002460:Cell Line; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli; D042822:Genomic Instability; D006801:Humans; D033862:Iron-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D016296:Mutagenesis; D065636:Myotonin-Protein Kinase; D009690:Nucleic Acid Conformation; D010957:Plasmids; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics*; D017421:Sequence Analysis; D020079:Terminal Repeat Sequences; D000096923:Frataxin
D004247:DNA; D033862:Iron-Binding Proteins; D065636:Myotonin-Protein Kinase; D017346:Protein Serine-Threonine Kinases; D000096923:Frataxin; C585517:DMPK protein, human