CAREER: The Role of Ribosome Conformational Dynamics on the Mechanism of Protein Synthesis

Columbia University, New York NY

Investigators

Abstract

This project focuses on one of the most fundamental of cellular processes: protein synthesis by the ribosome, a universally conserved ribonucleoprotein enzyme. The ultimate goal in the Gonzalez laboratory is to characterize the conformational dynamics of the ribosome and other biomolecules involved in protein synthesis and utilize these data to elucidate the detailed molecular mechanism of protein synthesis. Novel fluorescence spectroscopic approaches are used to study the ribosomal molecular machine at the single-molecule level, focusing on conformationally dynamic regions of the biomolecule whose functional importance remains largely mysterious. Using single-molecule fluorescence, the mechanics of conformational changes within the ribosome are identified, quantified and coupled to biochemical steps in the overall process of protein synthesis. Achieving this level of resolution, however, requires merging the areas of molecular biology, biochemistry, biophysics, optics, microscopy, surface chemistry, microfluidics, and statistical data analysis. The laboratory is therefore highly interdisciplinary and is actively engaged in complementary technological developments, such as the application of microfluidic design principles to the sample cells used for single-molecule fluorescence measurements and the development of mathematically robust statistical models for data analysis that will expand the capabilities of single-molecule fluorescence spectroscopies. Beyond exclusive impact in the area of protein synthesis, the general principles developed by this research are directly applicable to the study of the numerous complex biochemical reactions that are of critical functional importance to the cell. The interdisciplinary nature of the research in the Gonzalez laboratory provides a unique opportunity to integrate students of varied scientific backgrounds and stimulate cross-discipline training. In order to achieve similar educational impact in the broader departmental and university communities at Columbia University, Prof. Gonzalez is integrating nucleic acid and single-molecule biophysics into the curriculum, participating in the design of a shared molecular biology facility, and organizing a discussion forum for users of this facility. Prof. Gonzalez has also put in place a long-term undergraduate/high-school student research program composed of projects to be exclusively executed by undergraduate and high-school student researchers. This educational and practical experience in all aspects of an interdisciplinary research project will prepare students for future work in contemporary scientific laboratory settings. This project is cofunded by the Divisions of Molecular and Cellular Biology and Chemistry.