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Journal of Bacteriology, August 2003, p. 4609-4614, Vol. 185, No. 15
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.15.4609-4614.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification of Upstream Sequences Essential for Activation of a Bacteriophage P2 Late Promoter

Gail E. Christie,1* Douglas L. Anders,1,{dagger} Victor McAlister,1 Tina S. Goodwin,1 Bryan Julien,2,{ddagger} and Richard Calendar2

Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, Virginia,1 Department of Molecular and Cell Biology, University of California, Berkeley, California2

Received 25 July 2002/ Accepted 9 May 2003

We have carried out a mutational scan of the upstream region of the bacteriophage P2 FETUD late operon promoter, PF, 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.

* Corresponding author. Mailing address: P.O. Box 980678, Richmond, VA 23298-0678. Phone: (804) 828-9093. Fax: (804) 828-9946. E-mail: christie{at}hsc.vcu.edu.

{dagger} Present address: Hazardous Materials Response Unit, FBI Academy, Quantico, VA 22135.

{ddagger} Present address: Kosan Biosciences, Hayward, CA 94545.

Journal of Bacteriology, August 2003, p. 4609-4614, Vol. 185, No. 15
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.15.4609-4614.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • McAlister, V. J., Christie, G. E. (2009). Analysis of DNA Binding by a Eubacterial Zinc Finger Transcription Factor. J. Bacteriol. 191: 4513-4521 [Abstract] [Full Text]  
  • Markov, D., Christie, G. E., Sauer, B., Calendar, R., Park, T., Young, R., Severinov, K. (2004). P2 Growth Restriction on an rpoC Mutant Is Suppressed by Alleles of the Rz1 Homolog lysC. J. Bacteriol. 186: 4628-4637 [Abstract] [Full Text]  


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