PT  - JOURNAL ARTICLE
AU  - Debora Tenenbaum
AU  - Koe Inlow
AU  - Larry Friedman
AU  - Anthony Cai
AU  - Jeff Gelles
AU  - Jane Kondev
TI  - RNA polymerase sliding on DNA can couple the transcription of nearby bacterial operons
AID  - 10.1101/2023.02.10.528045
DP  - 2023 Jan 01
TA  - bioRxiv
PG  - 2023.02.10.528045
4099  - http://biorxiv.org/content/early/2023/02/10/2023.02.10.528045.short
4100  - http://biorxiv.org/content/early/2023/02/10/2023.02.10.528045.full
AB  - DNA transcription initiates after an RNA polymerase (RNAP) molecule binds to the promoter of a gene. In bacteria, the canonical picture is that RNAP comes from the cytoplasmic pool of freely diffusing RNAP molecules. Recent experiments suggest the possible existence of a separate pool of polymerases, competent for initiation, which freely slide on the DNA after having terminated one round of transcription. Promoter-dependent transcription reinitiation from this pool of post-termination RNAP may lead to coupled initiation at nearby operons, but it is unclear whether this can occur over the distance- and time-scales needed for it to function widely on a bacterial genome in vivo. Here, we mathematically model the hypothesized reinitiation mechanism as a diffusion-to-capture process and compute the distances over which significant inter-operon coupling can occur and the time required. These quantities depend on previously uncharacterized molecular association and dissociation rate constants between DNA, RNAP and the transcription initiation factor σ70; we measure these rate constants using single-molecule experiments in vitro. Our combined theory/experimental results demonstrate that efficient coupling can occur at physiologically relevant σ70 concentrations and on timescales appropriate for transcript synthesis. Coupling is efficient over terminator-promoter distances up to ∼ 1, 000 bp, which includes the majority of terminator-promoter nearest neighbor pairs in the E. coli genome. The results suggest a generalized mechanism that couples the transcription of nearby operons and breaks the paradigm that each binding of RNAP to DNA can produce at most one messenger RNA.SIGNIFICANCE STATEMENT After transcribing an operon, a bacterial RNA polymerase can stay bound to DNA, slide along it, and reini-tiate transcription of the same or a different operon. Quantitative single-molecule biophysics experiments combined with mathematical theory demonstrate that this reinitiation process can be quick and efficient over gene spacings typical of a bacterial genome. Reinitiation may provide a mechanism to orchestrate the transcriptional activities of groups of nearby operons.Competing Interest StatementThe authors have declared no competing interest.