Molecular Biology of Proteolysis in T4-Infected Cells

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This proposal focuses on the molecular biology of protein degradation in phage T4-infected Escherichia coli bacteria. The specific aims of this proposal include (a) the identification of T4 genes in addition to pinA that function to block E. coli proteolytic activities. (b) The characterization of T4 mutants that fail to block protein degradation. The methodology involves standard molecular biological and biochemical procedures and, in addition, novel screening and selection strategies based on the degradation of specific proteins. This research should lead to a broader knowledge of virus - host cell interactions, to an increased understanding of the molecular bases by which E. coli proteases recognize their substrates, and to an ability to reduce proteolysis in E. coli and, thereby, to stabilize proteins of medical and scientific importance produced in E. coli bacteria.