SGER: Strategies for Detailed Characterization of E. coli Ribosome Biogenesis

University Of Rochester, Rochester NY

Investigators

Abstract

One of the most pressing and important challenges in contemporary molecular biology is to understand how large macromolecular complexes are assembled in vivo. Of particular interest is the assembly of the ribosome. Ribosomes are large machines whose appropriate formation is critical for cell viability. The goal of this project is to develop new technologies to study ribosome assembly in vivo. These new strategies will allow determination of the temporal order of addition of ribosomal proteins (r-proteins) to the assembling complexes and to relate ribosomal RNA (rRNA) processing events to stages in this ordered pathway. This work will provide a set of novel tools and protocols for analyzing ribosomal subunit assembly in vivo and dissecting the role(s) of non-ribosomal factors in this process. This exploratory project will create entirely new strategies for studying macromolecular assembly. It is of great importance because of the immediate impact of this technology on the study and understanding of biogenesis of macromolecular complexes. The project will develop two, complementary technologies for the study of ribosome subunit assembly in vivo. Success in developing these strategies will fill an experimental void and allow detailed analysis of the necessary and required events for functional ribosome formation in the cell. Broader Impacts The project will allow training of a graduate student in state-of-the-art technologies and technology development. Additionally, once these strategies have proven successful in studies of ribosome assembly, they should be directly applicable to the study of other complex cellular machines.