title: Detection of RNA-DNA association by a proximity ligation-based method.
abstract: We describe a proximity ligation assay (PLA)-based method of assessing association of DNA and RNA in single cells during the cell cycle. Pulse-labeling of DNA with EdU and RNA with BrU and testing their close proximity by PLA demonstrates that RNA synthesis in individual cells resumes about 30-45 min after DNA replication. Consistent with this conclusion, RNA Pol II phosphorylated at Ser2 of its CTD is detected at the same time as RNA transcripts on nascent DNA. Our results also show that RNA is associated with DNA foci during all stages of mitosis.
authors.affiliation: Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Petruk
authors.affiliation: Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Tyler K
authors.initials: TK
authors.lastname: Fenstermaker
authors.affiliation: Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Black
authors.affiliation: Department of Zoology, University of British Columbia, 6270 University Boulevard, Vancouver, BC V6T 1Z4, Canada.
authors.forename: Hugh W
authors.initials: HW
authors.lastname: Brock
authors.affiliation: Department of Biochemistry and Molecular Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Mazo
mesh_terms: D002453:Cell Cycle; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008967:Molecular Biology / Q000379:methods; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000235:genetics* / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors
chemical_list: D012313:RNA; D004247:DNA