Michael S. Kilberg, CURRENT RESEARCH

TRANSCRIPTIONAL CONTROL OF THE HUMAN ASPARAGINE SYNTHETASE GENE

    Nutrient control of cellular processes has been well documented in bacteria and lower eukaryotic cells, but our knowledge in mammalian cells is relatively limited.  Our laboratory, as well as others, has identified several proteins for which synthesis is increased in response to amino acid deprivation of tissue or cells in culture.  For example, human asparagine synthetase (ASNS), the enzyme that catalyzes asparagine biosynthesis, is transcriptionally regulated enhanced following amino acid deprivation.  While the induction of ASNS activity following amino acid deprivation is well documented, the mechanisms responsible are not understood.  Our hypothesis is that the human asparagine synthetase gene represents a model system for investigating metabolite control of gene transcription.  The goal of this program is to characterize the cis-acting sequences within the human ASNS gene that are responsible for amino acid-dependent transcriptional control and to identify and characterize the trans-acting factors that bind to these sequences.

Students working on these projects will become familiar with techniques such as analysis of chromatin structure, in vivo footprinting, site-directed mutagenesis, promoter analysis, electrophoresis mobility shift assays (EMSA), chromatin immunoprecipitation (ChIP), retroviral and plasmid-based gene delivery, quantitative real-time PCR, and a wide range of other common molecular biology techniques.