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- Title
The bacteriophage T4 middle promoter P<sub>uvsX</sub>: analysis of regions important for binding of the T4 transcriptional activator MotA and for activation of transcription.
- Authors
March-Amegadzie, Roslyn; Hinton, Deborah M.
- Abstract
Bacteriophage T4 middle promoters, which are transcribed using phage-modified host RNA polymerase and the T4 transcriptional activator, MotA, match the host σ70 consensus sequence at -10, but they have a different consensus ((t/a)(t/a)TGCTT(t/c)A) (a MotA box) at -30. While the T4 middle promoter Puvsx has these -10 and -30 motifs, it also has matches to the MotA box at -35, -51, -70, and -87. We show that MotA binds to Puvsx DNA, footprinting a region that includes the MotA boxes at -30, -35, and -51. Very high levels of MotA are required for footprinting and gel-shift experiments, and protein-DNA complexes formed in the presence of both phage-modified polymerase and MotA are more resistant to <em>Hin</em>dlll cleavage than those formed with either protein alone. These results suggest that MotA-DNA interactions may be stabilized by phage-modified polymerase. Sequences between -18 and -38 are absolutely required for MotA activation of transcription, but sequences upstream of -38 are stimulatory, particularly when chloride instead of glutamate is the major anion. Our results dissect Puvsx into a core promoter, downstream of -38, which is required for MotA activation, and an upstream region that enhances transcription especially under conditions less favourable for protein-DNA interactions.
- Subjects
BACTERIOPHAGE T4; PROMOTERS (Genetics); GENETIC transcription; RNA polymerases; DNA
- Publication
Molecular Microbiology, 1995, Vol 15, Issue 4, p649
- ISSN
0950-382X
- Publication type
Article
- DOI
10.1111/j.1365-2958.1995.tb02374.x