title: Analysis of chromatin-nuclear receptor interactions by laser-chromatin immunoprecipitation.
abstract: Better defining the dynamics of biomolecular interactions is an important step in understanding molecular biology and cellular processes. DNA-protein interactions, and specifically hormone-triggered DNA-nuclear receptor interactions, are key events which are still poorly understood. To date, the most commonly used approach in studying chromatin interactions is the immunoprecipitation of chemically cross-linked chromatin (ChIP) coupled with single gene or global genomic analyses. Currently, establishing a stable interplay between nucleic acids and proteins (DNA-protein cross-link) is mainly obtained through conventional, diffusion-triggered, chemical methods using formaldehyde. Here we describe an alternative method, called Laser-ChIP (LChIP), for the specific analysis of interactions between chromatin and nuclear receptors driven by a UV laser energy source. Photo-induced cross-linking in LChIP is achieved very rapidly, allowing the study of transient interactions, depending on laser source parameters.
authors.affiliation: Dipartimento di Biochimica, Biofisica e Patologia Generale-II Università di Napoli, Via L. De Crecchio, 7, 80138, Naples, Italy.
authors.forename: Rosaria
authors.initials: R
authors.lastname: Benedetti
authors.forename: Mariarosaria
authors.initials: M
authors.lastname: Conte
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Carafa
authors.forename: Bartolomeo
authors.initials: B
authors.lastname: Della Ventura
authors.forename: Carlo
authors.initials: C
authors.lastname: Altucci
authors.forename: Raffaele
authors.initials: R
authors.lastname: Velotta
authors.forename: Hendrik G
authors.initials: HG
authors.lastname: Stunnenberg
authors.forename: Lucia
authors.initials: L
authors.lastname: Altucci
authors.forename: Angela
authors.initials: A
authors.lastname: Nebbioso
mesh_terms: D000818:Animals; D002453:Cell Cycle; D016923:Cell Death; D002843:Chromatin / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation / Q000295:instrumentation* / Q000379:methods; D004247:DNA / Q000378:metabolism; D004867:Equipment Design; D006801:Humans; D007834:Lasers; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*
chemical_list: D002843:Chromatin; D018160:Receptors, Cytoplasmic and Nuclear; D004247:DNA