Chemical Synthesis and Biological Studies on Mutated Gene-control Regions

Abstract

Many biochemically important processes are dependent on the interaction of various proteins with defined sequences of DNA. Some examples are replication, transcription, certain forms of recombination, positive and negative control of gene expression, and host restriction-modification enzymes. However, the mechanisms whereby these proteins recognize and bind to unique polynucleotide sequences are poorly understood. A major emphasis of our research is to understand these recognition processes. Our approach is to manipulate and modify specifically a gene-control region by chemical synthesis and then to study how these sequence-altered DNAs interact with the appropriate proteins. In the past, our research has focused on the lac repressor-lac operator (Caruthers 1980) and the cI repressor-λ operator (Kawashima et al. 1977) systems. Currently, we are also examining the interaction of SV40 T antigen, Escherichia coli RNA polymerase, cAMP receptor protein (CRP), and cro repressor with appropriate gene-control regions. In this paper we outline our recent results...