title: Identification of upstream sequences essential for activation of a bacteriophage P2 late promoter.
abstract: We have carried out a mutational scan of the upstream region of the bacteriophage P2 FETUD late operon promoter, P(F), which spans an element of hyphenated dyad symmetry that is conserved among all six of the P2 and P4 late promoters. All mutants were assayed for activation by P4 delta in vivo, by using a lacZ reporter plasmid, and a subset of mutants was assayed in vitro for delta binding. The results confirm the critical role of the three complementary nucleotides in each half site of the upstream element for transcription factor binding and for activation of transcription. A trinucleotide DNA recognition site is consistent with a model in which these transcription factors bind via a zinc finger motif. The mutational scan also led to identification of the -35 region of the promoter. Introduction of a sigma(70) -35 consensus sequence resulted in increased constitutive expression, which could be further stimulated by delta. These results indicate that activator binding to the upstream region of P2 late promoters compensates in part for poor sigma(70) contacts and helps to recruit RNA polymerase holoenzyme.
authors.affiliation: Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, Virginia, USA. christie@hsc.vcu.edu
authors.forename: Gail E
authors.initials: GE
authors.lastname: Christie
authors.forename: Douglas L
authors.initials: DL
authors.lastname: Anders
authors.forename: Victor
authors.initials: V
authors.lastname: McAlister
authors.forename: Tina S
authors.initials: TS
authors.lastname: Goodwin
authors.forename: Bryan
authors.initials: B
authors.lastname: Julien
authors.forename: Richard
authors.initials: R
authors.lastname: Calendar
mesh_terms: D017901:Bacteriophage P2 / Q000235:genetics*; D001483:Base Sequence; D015967:Gene Expression Regulation, Viral; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D014764:Viral Proteins