Compromised factor-dependent transcription termination in a nusA mutant of Escherichia coli : spectrum of termination efficiencies generated by perturbations of Rho, NusG, NusA, and the H-NS family of proteins

Abstract

The proteins NusA and NusG, which are essential for viability of wild-type Escherichia coli, participate in various post-initiation steps of transcription including elongation, antitermination, and termination. NusG is required along with the essential Rho protein for factor-dependent transcription termination (also referred to as polarity), but the role of NusA is less clear with conflicting reports that it both promotes and inhibits the process. In this study, we have found that a recessive missense nusA mutant (-R258C) exhibits a transcription termination-defective (that is, polarity-relieved) phenotype, much like missense mutants in rho or nusG, but is unaffected for either the rate of transcription elongation or antitermination in λ phage. Various combinations of the rho, nusG, and nusA mutations were synthetically lethal, and the lethality was suppressed by expression of the N-terminal half of nucleoid protein H-NS. Our results suggest that NusA function is indeed needed for factor-dependent transcription termination and that an entire spectrum of termination efficiencies can be generated by perturbations of the Rho, NusG, NusA, and H-NS family of proteins, with the corresponding phenotypes extending from polarity through polarity relief to lethality.