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. 2020 Sep 14;48(18):10353–10367. doi: 10.1093/nar/gkaa741

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 2. — Analysis of collisions between RNAP-template complexes and replisomes. (A) Cartoon illustrating the potential products of a collision between a replisome and a CO-RNAP-template complex. (B) CO RNAP-replication collisions cause transient fork stalling, whereas HO RNAP-replication collisions are severe blockages to fork progression. Time courses of replisome collisions on templates carrying either no RNAP complexes (on template CO₁₀₀, lanes 1–4); three different CO RNAP-templates: 19mer (CO₁₀₀), lanes 5–8, 100mer (CO₁₀₀), lanes 9–12, or RNAP array (CO₁₀₀), lanes 13–16; or three different HO RNAP-templates: 19mer (HO₁₀₀), lanes 17–20, 100mer (HO₁₀₀), lanes 21–24, or RNAP array (HO₁₀₀), lanes 25–28, analyzed by either (i) native agarose gel or (ii) denaturing agarose gel electrophoresis (n = 5). (C) Fork blockage becomes more severe as the complexity of the transcription complex increases. The fraction of stalled fork DNA products from native gels are plotted as a function of time for CO (i) and HO (ii) collisions. (iii) Reciprocal relationship between the fraction of stalled leading strand product and full-length material as a product of time for HO collisions on denaturing gels. (D) Relative efficiency of the DNA helicases Rep (blue) and UvrD (red) in resolving stalled forks in HO collisions with a 19mer RNAP (•, on template HO₁₉) or an RNAP array (▴, HO₁₀₀) (n = 3). SF, stalled fork; FL, full length product; RS, restart product; S, stalled leading-strand product; OF, Okazaki fragments.