---
title: "RfaH enables RNA polymerase processivity for long-range transcription"
id: "biorxiv-0-rfah-licenses-rna-polymerase-for-long-range-transcription"
canonical_url: "https://medichelpline.com/clinical-feed/biorxiv-0-rfah-licenses-rna-polymerase-for-long-range-transcription"
content_type: "clinical_feed_article"
specialty: "General"
source_name: "bioRxiv (Biomedical Preprints)"
source_url: "https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1?rss=1"
published_at: "2026-09-19T12:00:00.000Z"
evidence_level: "Verified Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# RfaH enables RNA polymerase processivity for long-range transcription
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/biorxiv-0-rfah-licenses-rna-polymerase-for-long-range-transcription
- **Specialty:** [General](https://medichelpline.com/clinical-feed/general.md)
- **Primary Source:** bioRxiv (Biomedical Preprints)
- **Source URL:** [Original Journal Publication](https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1?rss=1)
- **Published At:** 2026-09-19T12:00:00.000Z
- **Evidence Rating:** Verified Feed
## Executive GIST (TL;DR)
- The study examines how **RfaH**, a NusG paralog, promotes long-range transcription by Escherichia coli **RNA polymerase (RNAP)**, which must transcribe operons exceeding **10+ kbp** despite threats from long-lived, backtrack-induced pauses. - Using high-throughput magnetic tweezers, the authors reconstituted **ops**-induced pausing in vitro and showed that the local **sequence context** determines pause occupancy. - **RfaH** is recruited to **ops**-paused RNAP via two distinct loading pathways: one that allows immediate escape from the pause and another that requires rescue by the cleavage factor **GreA**. - Both **NusG** and **RfaH** counteract the pause-stimulating activity of **NusA**, with **RfaH** sustaining transcription runs approximately four times longer than NusG in the assays described. - The in vitro behavior of RfaH parallels its in vivo role: depletion of RfaH impairs expression of long virulence operons and leads to failure of conjugation, indicating biological relevance. - The study contrasts different factors that influence RNAP pausing and rescue: **Gre** factors (stimulate transcript cleavage), **NusA** (stimulates pausing), and **NusG** (suppresses pausing and bridges the expressome). - The work establishes an experimental platform and mechanistic framework for dissecting expression of long virulence operons and suggests possible points for therapeutic targeting, while noting this is a preprint not yet peer-reviewed.
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Luca Buccolieri 1 VU Amsterdam; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Luca%2BBuccolieri%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Buccolieri%20L&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ALuca%2BBuccolieri%2B) Bing Wang 2 The Ohio State University * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Bing%2BWang%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Wang%20B&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ABing%2BWang%2B) Pim America 1 VU Amsterdam; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Pim%2BAmerica%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=America%20P&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3APim%2BAmerica%2B) Irina Artsimovitch 2 The Ohio State University * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Irina%2BArtsimovitch%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Artsimovitch%20I&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3AIrina%2BArtsimovitch%2B) David Dulin 1 VU Amsterdam; * [Find this author on Google Scholar](https://www.biorxiv.org/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=David%2BDulin%2B "Open in new tab") * [Find this author on PubMed](https://www.biorxiv.org/lookup/external-ref?access_num=Dulin%20D&link_type=AUTHORSEARCH "Open in new tab") * [Search for this author on this site](https://www.biorxiv.org/search/author1%3ADavid%2BDulin%2B) * [ORCID record for David Dulin](http://orcid.org/0000-0003-4209-0377 "Open in new tab") * For correspondence: d.dulin@vu.nl * [Abstract](https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_art/node:5795708/1) * [Info/History](https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1.article-info)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_info/node:5795708/1) * [Metrics](https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1.article-metrics)[](https://www.biorxiv.org/panels_ajax_tab/article_tab_metrics/node:5795708/1) * [Supplementary material](https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1.supplementary-material)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_data/node:5795708/1) * [ Preview PDF](https://www.biorxiv.org/content/10.64898/2026.09.18.752654v1.full.pdf+html)[](https://www.biorxiv.org/panels_ajax_tab/biorxiv_tab_pdf/node:5795708/1) ![Loading](https://www.biorxiv.org/sites/all/modules/contrib/panels_ajax_tab/images/loading.gif) ## Abstract Processivity is essential for gene expression: Escherichia coli RNA polymerase (RNAP) must transcribe 10+ kbp operons without failure, yet backtrack-induced long-lived pauses threaten premature termination. Gre factors stimulate transcript cleavage to rescue backtracked RNAP, whereas NusA and NusG, which bridge the expressome, respectively stimulate and suppress pausing. How they cooperate to secure full-length transcription is unclear. Using high-throughput magnetic tweezers, we reconstitute ops-induced pausing, showing that sequence context sets pause occupancy. RfaH, a NusG paralog recruited at ops and essential for long virulence operons, loads onto ops-paused RNAP by two pathways: one permitting immediate escape, the other requiring GreA rescue. We find that both NusG and RfaH nullify NusA's pause-stimulating effect, RfaH sustaining runs four times longer than NusG and replicating its role in vivo, where RfaH depletion leads to conjugation failing. This work provides the foundation for dissecting virulence operon expression and its therapeutic targeting. ### Competing Interest Statement The authors have declared no competing interest. ## Funder Information Declared National Institute of General Medical Sciences, R01 GM067153 Netherlands Ministry of Education, Culture and Science (OCW) and the Netherlands Organisation for Scientific Research (NWO), 024.003.019 Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a [CC-BY-NC 4.0 International license](http://creativecommons.org/licenses/by-nc/4.0/). bioRxiv and medRxiv thank the following for their generous financial support: > The Chan Zuckerberg Initiative, Cold Spring Harbor Laboratory, the Sergey Brin Family Foundation, California Institute of Technology, Centre National de la Recherche Scientifique, Fred Hutchinson Cancer Center, Imperial College London, Massachusetts Institute of Technology, Stanford University, The University of Edinburgh, University of Washington, and Vrije Universiteit Amsterdam. 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