Figure 3.
The deletion of ARS305 reverses the orientation-dependent behaviour of CEB1. (A) Mode of CEB1 replication upon deletion of ARS305. CEB1 is replicated by forks emanating from ARS306. In orientation I, the G-quadruplex-forming strand is the template of lagging-strand replication whereas in orientation II, it is the template of leading-strand replication. This is the opposite of Figure 1C where CEB1 was replicated by the nearby ARS305. (B) Southern blot analysis of CEB1-1.8 behaviour placed in orientation I or II in WT cells (ORT6165-2 and ORT6166-52, respectively) treated or not with 10 μM of Phen-DC3, and in pif1Δ cells (ORT6169-2 and ORT6170-8, respectively). Genomic DNA was digested with ApaI/XhoI (orientation I) and ApaI/NcoI (orientation II). Other legends as in Figure 1D.