| Autor: |
Mueller AU; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065., Chen J; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065., Wu M; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065., Chiu C; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065., Nixon BT; Department of Biochemistry and Molecular Biology, Penn State University, University Park, PA 16802., Campbell EA; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065., Darst SA; Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY 10065. |
| Abstrakt: |
Bacterial transcription initiation requires σ factors for nucleation of the transcription bubble. The canonical housekeeping σ factor, σ 70 , nucleates DNA melting via recognition of conserved bases of the promoter -10 motif, which are unstacked and captured in pockets of σ 70 . By contrast, the mechanism of transcription bubble nucleation and formation during the unrelated σ N -mediated transcription initiation is poorly understood. Herein, we combine structural and biochemical approaches to establish that σ N , like σ 70 , captures a flipped, unstacked base in a pocket formed between its N-terminal region I (RI) and extra-long helix features. Strikingly, RI inserts into the nascent bubble to stabilize the nucleated bubble prior to engagement of the obligate ATPase activator. Our data suggest a general paradigm of transcription initiation that requires σ factors to nucleate an early melted intermediate prior to productive RNA synthesis. |
