title: Simultaneous single-molecule detection of endogenous C-5 DNA methylation and chromatin accessibility using MAPit.
abstract: Bisulfite genomic sequencing provides a single-molecule view of cytosine methylation states. After deamination, each cloned molecule contains a record of methylation within its sequence. The full power of this technique is harnessed by treating nuclei with an exogenous DNMT prior to DNA extraction. This exogenous methylation marks regions of accessibility and footprints nucleosomes, as well as other DNA-binding proteins. Thus, each cloned molecule records not only the endogenous methylation present (at CG sites, in mammals), but also the exogenous (GC, when using the Chlorella virus protein M.CviPI). We term this technique MAPit, methylation accessibility protocol for individual templates.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida and Shands Cancer Center, University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Russell P
authors.initials: RP
authors.lastname: Darst
authors.forename: Carolina E
authors.initials: CE
authors.lastname: Pardo
authors.forename: Santhi
authors.initials: S
authors.lastname: Pondugula
authors.forename: Vamsi K
authors.initials: VK
authors.lastname: Gangaraju
authors.forename: Nancy H
authors.initials: NH
authors.lastname: Nabilsi
authors.forename: Blaine
authors.initials: B
authors.lastname: Bartholomew
authors.forename: Michael P
authors.initials: MP
authors.lastname: Kladde
mesh_terms: D044503:5-Methylcytosine / Q000378:metabolism*; D001483:Base Sequence; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D003001:Cloning, Molecular; D019175:DNA Methylation / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D013447:Sulfites; D024363:Transcription Initiation Site
chemical_list: D002843:Chromatin; D013447:Sulfites; D044503:5-Methylcytosine; C042345:hydrogen sulfite