Chemical mapping of cytosines enzymatically flipped out of the DNA helix.
Nucleic Acids Res
; 36(10): e57, 2008 Jun.
Article
em En
| MEDLINE
| ID: mdl-18450817
ABSTRACT
Haloacetaldehydes can be employed for probing unpaired DNA structures involving cytosine and adenine residues. Using an enzyme that was structurally proven to flip its target cytosine out of the DNA helix, the HhaI DNA methyltransferase (M.HhaI), we demonstrate the suitability of the chloroacetaldehyde modification for mapping extrahelical (flipped-out) cytosine bases in protein-DNA complexes. The generality of this method was verified with two other DNA cytosine-5 methyltransferases, M.AluI and M.SssI, as well as with two restriction endonucleases, R.Ecl18kI and R.PspGI, which represent a novel class of base-flipping enzymes. Our results thus offer a simple and convenient laboratory tool for detection and mapping of flipped-out cytosines in protein-DNA complexes.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Enzimas de Restrição do DNA
/
Citosina
/
DNA (Citosina-5-)-Metiltransferases
Tipo de estudo:
Evaluation_studies
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article