Visualization of the binding site for the transcript cleavage factor GreB on Escherichia coli RNA polymerase.
The structure of Escherichia coli core RNA polymerase (RNAP) complexed with the transcript cleavage factor GreB was determined from electron micrographs of negatively stained, flattened helical crystals. A binding assay was developed to establish that GreB was incorporated into the RNA polymerase crystals with high occupancy through interactions between the globular C-terminal domain and the RNA polymerase. Comparison of the core RNAP:GreB structure with the previously determined structure of core RNAP located the GreB binding site on one face of the RNA polymerase, next to but not in the 25 A-diameter channel of RNA polymerase.
Laboratory of Molecular Biophysics, The Rockefeller University, 1230 York Avenue, New York, NY, 10021, USA.
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A
Polyakov
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Richter
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A
Malhotra
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D
Koulich
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Borukhov
S A
SA
Darst
D001665:Binding Sites; D002942:Circular Dichroism; D003460:Crystallization; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology*; D029968:Escherichia coli Proteins; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods; D014157:Transcription Factors / Q000737:chemistry*; D035602:Transcriptional Elongation Factors
D029968:Escherichia coli Proteins; C079668:GreA protein, E coli; C079667:GreB protein, E coli; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; D012321:DNA-Directed RNA Polymerases