title: RNA labeling and immobilization for nano-displacement measurement: probing three-dimensional RNA structure.
abstract: RNA molecules play essential roles in many biological processes, including the storage and transfer of information in the cell. These events are mediated via RNA-protein interactions or by catalytic RNA molecules. It is now recognized that unique RNA folds are associated with biological functions. Therefore, to study the intrinsic structural changes and dynamics which regulate the various functions of RNA, it is necessary to probe its three-dimensional structure in solution. In this respect, using single-molecule methodologies may allow study of native RNA molecules independent of their size and in real time. However, this may require the immobilization of RNA on a surface. Here, we report a novel approach to immobilize RNA on a glass. The procedures involve both chemical and enzymatic modifications of long RNA molecules. In addition, we demonstrate the application of an optical tweezers apparatus to measure the length and, hence, the dynamics of immobilized intact ribosomal RNA molecules as a function of different solution conditions.
authors.affiliation: Department of Structural Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Barak
authors.initials: B
authors.lastname: Akabayov
authors.forename: Arnon
authors.initials: A
authors.lastname: Henn
authors.forename: Michael
authors.initials: M
authors.lastname: Elbaum
authors.forename: Irit
authors.initials: I
authors.lastname: Sagi
mesh_terms: D000327:Adsorption; D005240:Feasibility Studies; D005898:Glass / Q000737:chemistry; D015636:Magnesium Chloride / Q000737:chemistry; D008846:Micromanipulation / Q000379:methods*; D009038:Motion; D036103:Nanotechnology / Q000379:methods*; D009690:Nucleic Acid Conformation; D055095:Optics and Photonics; D012313:RNA / Q000032:analysis / Q000737:chemistry* / Q000648:ultrastructure*; D012338:RNA, Ribosomal, 23S / Q000032:analysis / Q000737:chemistry / Q000648:ultrastructure; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D012338:RNA, Ribosomal, 23S; D015636:Magnesium Chloride; D012313:RNA