1. Analysis of a Double-Strand DNA Break in living S. cerevisiae Cells Leana M. Topper Kerry S. Bloom Department of Biology, University of North Carolina at Chapel Hill;

2. Chromosome III CEN MAT ~85 kb LacO array ~500 bp

3. 8 kb KpnI fragment HO cut fragment KpnI LacO MAT probe 2.5 kb No pGalHOT W/ pGalHOT 00.5 1 234 0 1 234 h post gal

4. Live cell after HO induction 2:00 6:007:00 15:00 8:00 10:0013:00

5. Movement of Spindle Pole Bodies and lacO in live cells following HO expression Average film time: 13 min No bud: 13 Small-budded cells: 8 Large-budded cells: 29 Average spindle length = 1.69 ±0.32  m Range = 1.05-2.44  m

6. Time post gal addition No budSmall budLarge bud 0 min8912 30 min3310 1 h289 2 h398 3 h856 4 h10614 Total 263147 Population analysis of lacO and SPBs after induction of HO

7. Deletion of Rad52 does not affect LacO movement 0:00 2:00 6:00 9:00 15:0020:00 21:00

8. Formation of Rad52 foci following DNA damage

9. Time post gal No bud foci Small bud foci Large bud foci Divided nucleus 0 h36013 3 (23%) 12 1 (8.3%) 9 1-2 h84023045 4 (8.8%) 21 2-3 h75022 2 (9.0%) 46 9 (19.6%) 13 3-4 h104 2 (1.9%) 18442 8 (19.0%) 21 Formation of Rad52-GFP foci after induction of HO

10. Rad52-GFP foci and Spindle Pole Bodies Move Independently 1:00 5:00 8:0012:00 17:0018:00 7:00

11. Rad52-CFP and LacO spots do not colocalize 3:00 8:006:00 9:00 11:0014:00 16:00