title: A PAS domain within F plasmid TraJ is critical for its function as a transcriptional activator.
abstract: TraJ is the positive activator of the major transfer operon in the F plasmid of Escherichia coli that counteracts H-NS silencing at the main transfer promoter (P(Y)). Multiple sequence alignment revealed a putative PAS (Per-ARNT-Sim) domain that might be involved in sensing redox potential or energy levels in the cell. This domain, which contains a conserved PXCXR motif along with a C(X)(9-10)CR/N/K motif of variable position, was identified within the N-terminal region of TraJ orthologues including F TraJ. The 5 cysteine residues in F TraJ were changed to serine to give protein with single or multiple substitutions. Single C to S substitutions had little effect on mating efficiency (ME), whereas cumulative substitutions from the N- to the C-termini (2CS to 5CS) gradually reduced ME to undetectable levels. F TraJ was able to bind to Fe (III) on an affinity sorbent column. This feature was severely impaired for the 5CS mutant. Thus, the cysteine residues within the PAS domain could be the part of a metal-containing redox centre that plays a key role in the transcriptional activation of the P(Y) operon by TraJ.
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Canada. arutyuno@ualberta.ca
authors.forename: D
authors.initials: D
authors.lastname: Arutyunov
authors.forename: J M
authors.initials: JM
authors.lastname: Rodriguez-Maillard
authors.forename: L S
authors.initials: LS
authors.lastname: Frost
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D002413:Cations, Divalent; D003545:Cysteine / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D005144:F Factor / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D002413:Cations, Divalent; D029968:Escherichia coli Proteins; C488999:TraJ protein, E coli; D014157:Transcription Factors; D003545:Cysteine