RIG: Integrated Research and Class Teaching on a Family of Microbial Transcription Factors

California State University-Fresno Foundation, Fresno CA

Investigators

Abstract

Intellectual Merit: Bacteria are everywhere on earth because they are masters at adapting to environmental changes. They sense their environment through specialized receptor proteins, such as the cAMP receptor protein (CRP) of Escherichia coli. This CRP protein has become the model for understanding transcriptional regulation in research and teaching. With many bacterial genomes sequenced in recent years, many CRP homologs have been discovered throughout the eubacteria. By analogy to E. coli CRP, these CRP homologs are predicted to be DNA binding proteins whose DNA binding activity is modulated by ligand binding. The main goal of this project is to apply the accumulated knowledge of the E. coli CRP to discover functions of these diverse CRP homologs. To identify the gene targets of the CRP homologs, the CRP homologs first will be mutated to introduce changes that should make them constitutively active; then the mutants will be used in SELEX (Systematic Evolution of Ligands by EXponential enrichment) experiments to identify their cognate high affinity target DNA sequences. A second strategy will identify ligands that influence activity of several CRP homologs and explore which amino acid residues are important for ligand recognition. Broader Impacts: The scientific goals of this project will be achieved using a format of integrated research and class teaching. A substantial portion of this research will be conducted by students within two lab-intensive courses: a molecular biology lab (~20 graduate students over two years, course offered once a year) and a microbiology lab (~30 undergraduate students over two years, course offered once a year). This hypothesis-driven project will give the students an excellent inquiry-based learning opportunity to experience how science is done and to consider science as a career option. Also, because approximately 40% of the students at California State University, Fresno are from underrepresented ethnic groups, this research project will have a significant impact on minority students.