96-00851 Flower Proteins destined for secretion are synthesized in precursor form with an amino terminal extension, the signal sequence. The latter is decoded by the protein export machinery which consists of sec proteins that are localized in the cytoplasm and in the membrane. Successful export of a protein requires sequential interactions among the various sec proteins. The long term goals of this research are to understand the functions of the sec proteins and to elucidate the requirements for productive interactions among the sec components. Escherichia coli is the model organism chosen for these studies. The translocation process in E. coli shares many similarities with the protein translocation event that is observed in the endoplasmic reticulum of higher organisms. The investigator will use a combination of genetic and biochemical methods to gain insights into protein export. The project has the following objectives: a) To use an in vitro system to study altered translocation processes in suppressor forms of SecE and SecY; b) To study SecA interactions with precursor proteins that have mutated signal sequences; and c) To generate synthetic lethal mutants of sec components to identify interacting domains. The educational component of this CAREER grant involves modernization of courses that are taught to students. To enhance comprehension of scientific concepts, teaching will also be performed within a laboratory. Also, mentoring of American Indian and Alaskan Natives will be made possible through Outreach programs. %%% Proteins destined for secretion are synthesized in precursor form with an amino terminal extension, the signal sequence. The latter is decoded by the protein export machinery which consists of sec proteins that are localized in the cytoplasm and in the membrane. Successful export of a protein requires sequential interactions among the various sec proteins. The long term goals of this research are to understand the functions of the sec proteins and to elucidate the requ irements for productive interactions among the sec components. Escherichia coli is the model organism chosen for these studies. The translocation process in E. coli shares many similarities with the protein translocation event that is observed in the endoplasmic reticulum of higher organisms. The investigator will use a combination of genetic and biochemical methods to gain insights into protein export. The educational component of this CAREER award involves modernization of courses that are taught to students. To enhance comprehension of scientific concepts, teaching will also be performed within a laboratory. Also, mentoring of American Indian and Alaskan Natives will be made possible through Outreach programs. ***
