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Science 23 May 1997:
Vol. 276. no. 5316, pp. 1258 - 1260
DOI: 10.1126/science.276.5316.1258
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Reports

Promoter Recognition As Measured by Binding of Polymerase to Nontemplate Strand Oligonucleotide

M. T. Marr and J. W. Roberts

In transcription initiation, the DNA strands must be separated to expose the template to RNA polymerase. As the closed initiation complex is converted to an open one, specific protein-DNA interactions involving bases of the nontemplate strand form and stabilize the promoter complex in the region of unwinding. Specific interaction between RNA polymerase and the promoter in Escherichia coli was detected and quantified as the binding affinity of nontemplate oligonucleotide sequences. The RNA polymerase subunit sigma factor 70 contacted the bases of the nontemplate DNA strand through its conserved region 2; a mutation that affected promoter function altered the binding affinity of the oligonucleotide to the enzyme.

Section of Biochemistry, Molecular and Cell Biology, Biotechnology Building, Cornell University, Ithaca, NY 14853, USA.

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C.A. GROSS, C. CHAN, A. DOMBROSKI, T. GRUBER, M. SHARP, J. TUPY, and B. YOUNG (1998)
Cold Spring Harb Symp Quant Biol 63, 141-156
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DNA Melting within a Binary sigma 54-Promoter DNA Complex.
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C. L. Chan and C. A. Gross (2001)
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M. Tomsic, L. Tsujikawa, G. Panaghie, Y. Wang, J. Azok, and P. L. deHaseth (2001)
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L. Milne, Y. Xu, D. M. Perrin, and D. S. Sigman (2000)
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Function of the bacterial TATAAT -10 element as single-stranded DNA during RNA polymerase isomerization.
M. S. Fenton and J. D. Gralla (2001)
PNAS 98, 9020-9025
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