title: A structural model for the assembly of the 30S subunit of the ribosome.
abstract: The order in which proteins bind to 16S rRNA, the assembly map, was determined by Nomura and co-workers in the early 1970s. The assembly map shows the dependencies of binding of successive proteins but fails to address the relationship of these dependencies to the three-dimensional folding of the ribosome. Here, using molecular mechanics techniques, we rationalize the order of protein binding in terms of ribosomal folding. We determined the specific contacts between the ribosomal proteins and 16S rRNA from a crystal structure of the 30S subunit (1FJG). We then used these contacts as restraints in a rigid body Monte-Carlo simulation with reduced-representation models of the RNA and proteins. Proteins were added sequentially to the RNA in the order that they appear in the assembly map. Our results show that proteins nucleate the folding of the head, platform, and body domains, but they do not strongly restrict the orientations of the domains relative to one another. We also examined the contributions of individual proteins to the formation of binding sites for sequential proteins in the assembly process. Binding sites for the primary binding proteins are generally more ordered in the naked RNA than those for other proteins. Furthermore, we examined one pathway in the assembly map and found that the addition of early binding proteins helps to organize the RNA around the binding sites of proteins that bind later. It appears that the order of assembly depends on the degree of pre-organization of each protein's binding site at a given stage of assembly, and the impact that the binding of each protein has on the organization of the remaining unoccupied binding sites.
authors.affiliation: Department of Biochemistry and Molecular Genetics, The University of Alabama at Birmingham, 1918 University Blvd, 552 Basic Health Sciences Bldg, Birmingham, AL 35294-0005, USA.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Stagg
authors.forename: Jason A
authors.initials: JA
authors.lastname: Mears
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Harvey
mesh_terms: D001665:Binding Sites; D003198:Computer Simulation; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D009010:Monte Carlo Method; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism*; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D012336:RNA, Ribosomal, 16S; D012269:Ribosomal Proteins