Figure 4: Replication fork breakage at a site-specific SSB. | Scientific Reports

Figure 4: Replication fork breakage at a site-specific SSB.

From: The RecQ DNA helicase Rqh1 constrains Exonuclease 1-dependent recombination at stalled replication forks

Figure 4

(a) Schematic of the ade6− direct repeat on chromosome 3 showing the position of the gpII nick site, probes, and relevant restriction sites and fragment sizes. Asterisks indicate the position of the point mutations in ade6-L469 and ade6-M375, and arrows show the direction of replication and transcription as indicated. The distance between the ade6− heteroalleles is ~5 kb. (b) Model for how replication forks are thought to break at SSBs in the leading or lagging template strand. (c) Alkaline and neutral gel analysis of genomic DNA from wild-type and rad50S strains containing either a bottom strand (BS) or top strand (TS) nick site, and either pREP1 or pREP1-NLS-gpII. DNA was digested with NdeI and detected with Probe A. For the rad50S strains, analysis of genomic DNA from two independent pREP1-NLS-gpII transformants is shown. (d) The same as (c) except Probe E is used instead of Probe A. (e) Neutral gel analysis of genomic DNA from exo1∆ rqh1∆ and rad50S strains containing either a BS or TS nick site, and either pREP1 or pREP1-NLS-gpII as indicated. DNA was digested with PvuII and SacI and detected with Probe A and B as indicated.

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