YidC-Structure, Function and Substrate Specificity

Ohio State University, The, Columbus OH

Investigators

Abstract

Intellectual Merit The long-term objective of this project is to understand how proteins are inserted into biological membranes. The majority of bacterial proteins are inserted into the membrane by the Sec translocase, although a subset is inserted and folded by the YidC translocase. The goal of the research project is to address the novel function of YidC in the generation of the tertiary structure of membrane proteins. The project has three important aims: (1) Determine the substrate specificity of YidC by identifying the features of a membrane protein that make it YidC dependent for insertion. (2) Elucidate the function of YidC in the insertion and folding of integral membrane proteins. (3) Define the transmembrane helix organization of the YidC monomer, and evaluate/extend the YidC dimer model. To achieve these aims, site-directed mutagenesis, YidC depletion strains, protein modification, cysteine crosslinking, and biochemical methods will be used. These studies will provide clues to similar insertion pathway in eukaryotic cells and provide important information in the biogenesis of membrane proteins such as receptors, channels, transporters, and ATPases. This research is significant in that it targets an important and unique protein, YidC, an evolutionarily conserved and essential protein involved in membrane protein biogenesis. The results of this project will enrich our understanding of the molecular machines involved in the insertion and folding of proteins and will be useful in understanding aberrant membrane protein folding. Broader Impact The laboratory is active in training and teaching undergraduate and graduate students. An important part of the undergraduate training is to motivate students to make discoveries and allow them to test hypotheses. The student's results from this project will be disseminated at conferences and published in top-tiered journals. The research on membrane protein insertion will be performed in a multidisciplinary research environment involving molecular biology and biochemical approaches. The lab will recruit and train minority students in research and encourage them to pursue graduate research in the life sciences area. Dr. Dalbey believes that an interest in the life sciences and research must be cultivated at an early age. He will pursue relationships with local schools K-12 to introduce the students to the excitement of scientific discovery.