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C700098200v1
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Papers In Press, published online ahead of print May 25, 2007
J. Biol. Chem, 10.1074/jbc.C700098200
Submitted on May 17, 2007
Revised on May 24, 2007
Accepted on May 24, 2007

Multi-subunit RNA polymerases melt only a single DNA base pair downstream of the active site

Ekaterina Kashkina, Michael Anikin, Florian Brueckner, Elisabeth Lehmann, Sergey N. Kochetkov, William T. McAllister, Patrick Cramer, and Dmitry Temiakov

Cell Biology, UMDNJ SOM, Stratford, NJ 08084

Corresponding Author: d.temiakov{at}umdnj.edu

To extend the nascent transcript, RNA polymerases must melt the DNA duplex downstream from the active site to expose the next acceptor base for substrate binding and incorporation. A number of mechanisms have been proposed to account for the manner in which the correct substrate is selected, and these differ in their predictions as to how far the downstream DNA is melted. Using fluorescence quenching experiments we provide evidence that cellular RNA polymerases from bacteria and yeast melt only one DNA base pair downstream from the active site. These data argue against a model in which multiple NTPs are lined up downstream of the active site.


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[Abstract] [Full Text] [PDF]


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