The Amino Terminus of Salmonella enterica Serovar Typhimurium ς 54 Is Required for Interactions with an Enhancer-Binding Protein and Binding to Fork Junction DNA

Abstract

ABSTRACT Transcription initiation by the ς 54 -RNA polymerase holoenzyme requires an enhancer-binding protein that is thought to contact ς 54 to activate transcription. To identify potential enhancer-binding protein contact sites in ς 54 , we compared the abilities of wild-type and truncated forms of Salmonella enterica serovar Typhimurium ς 54 to interact with the enhancer-binding protein DctD in a chemical cross-linking assay. Removal of two regions in the amino-terminal portion of ς 54 , residues 57 to 105 and residues 144 to 179, prevented cross-linking, but removal of either region alone did not. In addition, deletion of 56 amino-terminal residues of ς 54 (region I) reduced the affinity of the protein for a fork junction DNA probe.