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Science 27 February 2004:
Vol. 303. no. 5662, pp. 1382 - 1384
DOI: 10.1126/science.1092462
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Reports

Minimal Machinery of RNA Polymerase Holoenzyme Sufficient for Promoter Melting

Brian A. Young,* Tanja M. Gruber,* Carol A. Gross{dagger}

We determined the minimal portion of Escherichia coli RNA polymerase (RNAP) holoenzyme able to accomplish promoter melting, the crucial step in transcription initiation that provides RNAP access to the template strand. Upon duplex DNA binding, the N terminus of the ß' subunit (amino acids 1 to 314) and amino acids 94 to 507 of the {sigma} subunit, together comprising less than one-fifth of RNAP holoenzyme, were able to melt an extended –10 promoter in a reaction remarkably similar to that of authentic holoenzyme. Our results support the model that capture of nontemplate bases extruded from the DNA helix underlies the melting process.

Departments of Stomatology and Microbiology and Immunology, University of California, San Francisco, San Francisco, CA 94143, USA.



* These authors contributed equally to the work.

{dagger} To whom correspondence should be addressed. E-mail: cgross{at}cgl.ucsf.edu

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