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A general mechanism for transcription bubble nucleation in bacteria.


ABSTRACT: Bacterial transcription initiation requires σ factors for nucleation of the transcription bubble. The canonical housekeeping σ factor, σ70, nucleates DNA melting via recognition of conserved bases of the promoter -10 motif, which are unstacked and captured in pockets of σ70. By contrast, the mechanism of transcription bubble nucleation and formation during the unrelated σN-mediated transcription initiation is poorly understood. Herein, we combine structural and biochemical approaches to establish that σN, like σ70, 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.

SUBMITTER: Mueller AU 

PROVIDER: S-EPMC10083551 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature

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A general mechanism for transcription bubble nucleation in bacteria.

Mueller Andreas U AU   Chen James J   Wu Mengyu M   Chiu Courtney C   Nixon B Tracy BT   Campbell Elizabeth A EA   Darst Seth A SA  

Proceedings of the National Academy of Sciences of the United States of America 20230327 14


Bacterial transcription initiation requires σ factors for nucleation of the transcription bubble. The canonical housekeeping σ factor, σ<sup>70</sup>, nucleates DNA melting via recognition of conserved bases of the promoter -10 motif, which are unstacked and captured in pockets of σ<sup>70</sup>. By contrast, the mechanism of transcription bubble nucleation and formation during the unrelated σ<sup>N</sup>-mediated transcription initiation is poorly understood. Herein, we combine structural and b  ...[more]

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