CAREER: Kinetic Mechanism of DEAD-box RNA Helicase ATPase

Yale University, New Haven CT

Investigators

Abstract

DEAD-box proteins are ubiquitous enzymes that facilitate RNA structural rearrangements through the disruption of RNA-RNA and RNA-protein contacts in reactions that are coupled to the binding and hydrolysis of ATP. DEAD-box proteins are essential in all aspects of RNA metabolism including splicing, ribosome biogenesis, RNA degradation, and translation in all organisms from bacteria to humans. The E. coli DEAD-box protein DbpA is unique among the DEAD-box protein family in that it is specifically activated by the peptidyl-transferase center (PTC) RNA of the ribosome. Research efforts focus on providing a complete kinetic and thermodynamic characterization of the RNA-activated ATPase cycle of DbpA and defining how ATP utilization is coupled to PTC-RNA binding and unwinding. The detailed analysis of enzymatic reaction mechanisms will define how ATP utilization, RNA binding and RNA duplex unwinding are coupled, and advance our understanding of how DEAD box proteins use the energy from ATP hydrolysis to control the structure and dynamics of RNA. Knowledge acquired from the research activities will help develop testable hypotheses regarding the functions and molecular mechanisms of related DEAD-box RNA helicases. Broader impacts: The activities in this project will advance learning and education through graduate and undergraduate teaching and training, integrating chemistry with biology, and promoting the participation and contributions of underrepresented groups in the biomedical sciences. Undergraduate and graduate students will receive training in the methods of molecular biology, enzymology, and RNA and protein chemistry. A course on Enzyme Mechanisms will de developed further and include a comparison of ATPase reaction kinetics and energy transduction mechanisms of myosin and helicase molecular motors. The PI will continue to be active in underrepresented minority outreach activities, advising undergraduate and graduate students, and chairing the minority and graduate recruiting committees of the department.